CN101370818A - RNAi suppresses replication of influenza virus - Google Patents

Abstract

The invention relates to compositions and methods for modulating the expression of influenza viral genes, and more particularly to the downregulation of influenza viral genes by chemically modified oligonucleotides.

Description

RNAi suppresses replication of influenza virus

Cross References to Related Applications

This application claims priority over U.S. Application Serial Nos. 60/732,243, filed November 1, 2005, U.S. Serial Nos. 60/748,317, filed December 7, 2005, and U.S. Serial Nos. 60/799,000, filed May 9, 2006 right. The entire contents of each provisional application are hereby incorporated by reference.

technical field

The present invention relates to the field of compositions and methods for the treatment of influenza virus and the regulation of viral replication, in particular, the downregulation of influenza virus genes by means of RNA interference via oligonucleotides via inhalation/intranasal administration , topically to the lungs and nasal passages, or systemically, for example by intravenous injection.

Background of the invention

RNA interference or RNAi is a term originally coined by Fire and coworkers (Fire et al., Nature 391 :806-811, 1998) to describe the repression of gene expression when double-stranded RNA (dsRNA) is introduced into worms this observation. Short dsRNAs direct gene-specific post-transcriptional silencing in many organisms, including vertebrates, and provide new tools for studying gene function. There have been many recent reviews of this technique, see eg Novina, CD, and Sharp, P., Nature 2004, 430:161 and Sandy, P., et al., Biotechniques 2005, 39:215, which are incorporated herein by reference.

Influenza is one of the most widespread infections worldwide. It can be fatal: during the 1918 influenza A virus epidemic, an estimated 20-40 million people died. In the United States, approximately 20,000-40,000 people die each year from influenza A virus infection or its complications. During epidemics, flu-related hospitalizations can reach more than 300,000 in a single winter.

Several features contribute to the successful circulation of influenza viruses. First, it spreads easily between people through the air (droplet infection). Second, small changes in influenza virus antigens frequently occur (antigenic drift), such that the virus tends to evade protective immunity induced by previous infections with different virus variants. Third, new strains of influenza virus are readily generated by rearrangement or mixing of genetic material between different strains (antigenic shift). In the case of influenza A viruses, such mixing can occur between subtypes or strains that infect different species. The 1918 pandemic was said to have been caused by a viral hybrid strain derived from a combination between porcine and human influenza A viruses. Currently, there is widespread concern about the emergence of novel influenza strains potentially infecting humans, in particular influenza variants from avian origin, and more particularly from strain H5N1, through mixing among persons exposed to both human and avian influenza viruses. Experts familiar with most Asian societies and who have had close contact with agricultural birds and their keepers believe that it is not a question of if such mixed strains emerge, but only when. Worldwide epidemics worse than the one in 1918 may quickly follow.

Despite increased efforts, there is still no effective therapy for influenza virus infection, and the limited value of existing vaccines is due in part to the antigenic shift and drift described above. For these reasons, influenza A viruses have been under global surveillance for many years and are listed by the National Institutes of Health as one of the highest priority pathogens for biodefense. Although current vaccines based on inactivated viruses prevent disease in about 70-80% of healthy individuals under the age of 65, this percentage is much lower in older or immunocompromised individuals. Additionally, the costs and potential side effects associated with vaccine administration make this approach less than ideal. Although the antiviral drugs currently approved in the United States for the treatment and/or prevention of influenza are helpful, their use is limited due to concerns about side effects, compliance, and the possible emergence of resistant strains.

US Patent Application 20040242518 and corresponding WO 04/028471, both filed on September 29, 2003, recommend a small number of RNAi agents for the treatment of influenza. Their efficacy in humans has not been published.

Therefore, there remains a need to develop effective therapies for the treatment and prevention of viral infections in humans and animals, especially those that can efficiently target a wide range of viral subtypes. A prerequisite for high efficacy is that the active ingredient is not rapidly degraded in a physiological environment.

Summary of the invention

The present invention is based on in vitro and in vivo demonstration that influenza can be inhibited by intranasal administration of iRNA agents as well as by parenteral administration of such agents and identification of effective iRNA agents from the MP, NP, PB1, PB2 or PA genes of influenza virus For viral infection, the iRNA agent can reduce the RNA levels of many subtypes of influenza virus. Based on these findings, the present invention provides specific compositions and methods useful for reducing influenza virus mRNA levels, influenza virus protein levels, and viral titers of influenza virus in a subject, such as a mammal, eg, a human.

In particular, the present invention provides an iRNA agent consisting of, consisting essentially of or containing at least 15 or more of an influenza virus gene, in particular one of the influenza virus MP, NP, PB1, PB2 and PA genes Contiguous nucleotides, more specifically, reagents comprising 15 or more contiguous nucleotides from one of the sequences shown in Tables 1A-1H are provided. The iRNA agent preferably contains less than 30 nucleotides per strand, such as 21-23 nucleotides, as shown in Tables 1A-1H. Double-stranded iRNA reagents may have blunt ends or, more preferably, have a 1-4 nucleotide overhang from one or both 3' ends of the reagent.

Furthermore, iRNA agents may contain only naturally occurring ribonucleotide subunits, or may be synthetic so as to contain in the agent one or more modifications to the sugar or base of one or more ribonucleotide subunits. kind of modification. The iRNA agent can be further modified to attach it to a ligand, such as cholesterol, that selectively increases the stability, distribution, or cellular uptake of the agent. The iRNA agent may also be in isolated form, or be part of a pharmaceutical composition for use in the methods described herein, particularly as a pharmaceutical composition formulated for delivery to the lung or nasal cavity or for parenteral administration a part of. The pharmaceutical composition may contain one or more iRNA agents, and in some embodiments, two or more iRNA agents, each directed to a different sequence of an influenza virus gene or to a different influenza virus Gene.

In one aspect, the invention relates to double-stranded oligonucleotides comprising at least one unnatural nucleobase. In certain embodiments, the unnatural base is difluorotolyl, nitroindolyl, nitropyrrolyl, or nitroimidazolyl. In a preferred embodiment, the unnatural base is difluorotolyl. In certain embodiments, only one of the two oligonucleotide strands comprising a double stranded oligonucleotide contains an unnatural base. In certain embodiments, both oligonucleotide strands comprising double-stranded nucleotides independently contain unnatural bases.

The present invention also provides methods for reducing influenza virus viral RNA in cells. Such methods include the step of administering to a subject an iRNA agent of the invention as further described below. The method utilizes the cellular machinery associated with RNA interference that selectively degrades viral RNA in cells and includes the step of contacting the cells with an antiviral iRNA agent of the invention. The methods can be performed directly on cells, or can be performed on a mammalian subject by administering to the subject an iRNA agent/pharmaceutical composition of the invention. Reduction of viral RNA in cells leads to a reduction in the amount of viral protein produced and, in living organisms, to a reduction in replicating viral titer (as shown in the Examples).

The methods and compositions of the invention, eg, methods and iRNA agent compositions, can be used in any of the dosages and/or formulations set forth herein, and in any of the routes of administration set forth herein. Of particular importance, intranasal administration of iRNA agents and their ability to inhibit viral replication in respiratory tissues is shown herein.

The details of one or more embodiments of the invention are presented below along with illustrations and descriptions. Other features, objects and advantages of the invention will be apparent from the description, drawings and claims. This application incorporates by reference all cited documents, patents and patent applications in their entirety for all purposes.

Description of drawings

Figures 1A-1I: Dose-response curves of inhibition of target gene expression by selected RNAi agents. The respective target genes were recombinantly cloned in plasmids into Cos-7 cells, allowing the expression of mRNAs encoding the target genes and Renilla luciferase, the cells were treated with RNAi reagents, and quantified with Renilla luciferase. The cells were treated with RNAi reagents at concentrations of 100nM, 25nM, 6.3nM, 1.6nM, 400pM, 100pM, 24pM, 6pM, 1.5pM and 380fM, and by parametrized curve fitting, using the XLfit program, Determine _IC50 values.

Detailed description of the invention

The term "influenza virus" is used herein to refer to any strain of influenza virus that is capable of causing disease in animal or human subjects, or that is an interesting candidate for assay analysis. Influenza viruses are described in Fields, B., et al., Fields' Virology, 4 ^th ed. 2001, Lippincott Williams and Wilkins; Philadelphia, ISBN: 0781718325. In particular, the term includes any strain of influenza A virus that is capable of causing disease in animal or human subjects, or that is a candidate of interest for assay analysis. A large number of influenza A isolates have been fully or partially sequenced. Shown in Table 6 is only a partial list of the complete sequence of the influenza A genome fragment, which is stored in a public database (The influenza sequence database (The influenza Sequence Database, ISD)), referring to Macken, C., Lu, H., Goodman, J., & Boykin, L., "The value of adatabase in surveillance and vaccine selection." in Options for the Control of influenza IV. ADMEOsterhaus, N. Cox & AW Hampson (Eds.) 2001, Elsevier Science, Amsterdam, pp. 103- 106). This database also contains the complete sequences of the influenza B and C genome segments. The database is available on the Internet and includes a convenient search engine that allows users to search by genome segment, by type of viral infection, and by year of isolation. Influenza sequences are also available on GenBank. Accordingly, influenza gene sequences are readily available or readily determined by one of ordinary skill in the art.

For ease of description, the term "nucleotide" or "ribonucleotide" is sometimes used herein with reference to one or more monomeric subunits of an RNA agent. It should be understood that the use of the term "ribonucleotide" or "nucleotide" herein, in the case of a modified RNA or nucleotide substitute, also refers to a nucleotide modified at one or more positions Or the moiety substituted by a substitute, as further described below.

As used herein, "RNA reagent" refers to unmodified RNA, modified RNA, or nucleoside surrogates, each described herein or as known in the art of RNA synthesis. Although many modified RNAs and nucleoside surrogates have been described, preferred examples include those that are more resistant to nuclease degradation than unmodified RNAs. Preferred examples include having a 2' sugar modification, a single-stranded overhang (preferably a 3' single-stranded overhang), or, particularly if single-stranded, including one or more phosphate groups or one or more 5' modification of a phosphate analogue.

As used herein, an "iRNA agent" (short for interfering RNA agent) is an RNA agent that down-regulates the expression of a target gene, such as influenza virus. Without wishing to be bound by theory, iRNA agents may act by one or more of a number of mechanisms, including post-transcriptional cleavage of a target mRNA (sometimes referred to in the art as RNAi), or pre-transcriptional or pre-translational mechanisms. The iRNA agent can be a double-stranded iRNA agent.

As used herein, a "ds siRNA agent" (short for double-stranded iRNA agent) is an iRNA agent comprising more than one, preferably 2, strands, wherein hybridization between the strands can form regions of a duplex structure. As used herein, "strand" refers to a contiguous sequence of nucleotides, including non-naturally occurring or modified nucleotides. The two or more chains may be separate molecules, or each form part of a separate molecule, or they may be covalently linked to each other eg via a linker, eg a polyethylene glycol linker, to form one molecule. At least one strand may include a region that is substantially complementary to the target RNA. Such strands are termed "antisense strands". The second strand of the dsRNA agent, which contains the region that is complementary to the antisense strand, is termed the "sense strand." However, d siRNAs can also be formed from single RNA molecules that are at least partially self-complementary, forming, for example, hairpin or panhandle structures that include duplex regions. The latter are referred to herein as short hairpin RNAs (short hairpin RNAs) or shRNAs. In this context, the term "strand" refers to a region of an RNA molecule that is complementary to another region in the same RNA molecule.

While long ds iRNA agents can induce an often deleterious interferon response in mammalian cells, short ds iRNA agents do not trigger an interferon response, at least not to an extent that is deleterious to the cell and/or host (Manche et al., Mol. Cell. Biol. 12 :5238,1992; Lee et al., Virology 199 :491,1994; Castelli et al., J.Exp.Med.186 :967,1997; Zheng et al., RNA 10 :1934,2004; Heidel et al., "Lack of interferon response in animals to naked siRNAs"Nature Biotechn.advance online publication doi:10.1038/nbtl038, Nov.21, 2004). The iRNA agents of the invention include molecules that are short enough not to trigger deleterious non-specific interferon responses in normal mammalian cells. Thus, administering to a subject a composition comprising an iRNA agent (eg, formulated as described herein) can be used to reduce the expression of influenza virus genes in the subject's influenza virus expressing cells while circumventing an interferon response. Molecules that are short enough not to trigger a deleterious interferon response are referred to herein as siRNA agents or siRNAs. As used herein, "siRNA agent" or "siRNA" refers to an iRNA agent, such as a ds iRNA agent, which is short enough not to induce a deleterious interferon response in mammalian, particularly human, cells, e.g., which has less than 60 but preferably less than Duplex regions of 50, 40 or 30 nucleotide pairs.

As described herein, isolated iRNA agents, including ds siRNA agents and siRNA agents, can mediate decreased expression of influenza virus nucleic acid, e.g., by RNA degradation. For convenience, this RNA is also referred to herein as the RNA to be silenced. Such nucleic acids are also referred to as target genes. Preferably, the RNA to be silenced is the gene product of an influenza virus gene that is part of an influenza virus strain that is pathogenic to humans.

As used herein, the phrase "mediated RNAi" refers to the ability of an agent to silence a target gene in a sequence-specific manner. "Silencing a target gene" means the process whereby cells containing and/or expressing certain target gene products, when not exposed to an agent, contain and/or express at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% less said gene product than when contacted with the agent. The target gene product can be, for example, a messenger RNA (mRNA), protein or regulatory element.

As used herein, the term "complementary" is used to denote a sufficient degree of complementarity to allow stable and specific binding to occur between a compound of the invention and a target RNA molecule, such as influenza virus mRNA. Specific binding requires a sufficient degree of complementarity to avoid non-specific binding of non-target sequences and oligomeric compounds under conditions requiring specific binding, i.e., under physiological conditions in the context of in vivo assays or therapeutic treatments, or in In the case of an in vitro assay, under the conditions under which the assay is performed. Non-target sequences typically differ from the target sequence by at least 2, 3 or 4 nucleotides.

As used herein, an iRNA agent is "substantially complementary" to a target RNA, eg, a target mRNA (eg, target influenza virus mRNA), if the iRNA agent reduces production of a protein encoded by the target RNA in a cell. The iRNA agent can also be "precisely complementary" to the target RNA, eg, the target RNA and the iRNA agent anneal to form a hybrid consisting entirely of Watson-Crick base pairs, preferably at regions of exact complementarity. A "sufficiently complementary" iRNA agent can include an internal region (eg, an internal region of at least 10 nucleotides) that is exactly complementary to a target influenza virus RNA. Also, in some embodiments, iRNA agents are capable of specifically distinguishing single nucleotide differences. In this case, the iRNA agent only mediates RNAi if the exact complementarity is found at the region of a single nucleotide difference (eg, within 7 nucleotides). Preferred iRNA agents are based on, contain, or consist of the sense and antisense sequences provided in Tables 1A-1H.

As used herein, "substantially the same" when used to refer to a first nucleotide sequence compared with a second nucleotide sequence means that the first nucleotide sequence has at most 1 or 2 or 3 nucleotide substitutions (eg, uracil instead of adenosine) are identical to the second nucleotide sequence. As used herein, for an iRNA agent other than but derived from one of the iRNA agents in Tables 1A-1H by deletion, addition, or substitution of nucleotides, "substantially remains in cultured human influenza virus expression The ability to inhibit expression of influenza virus in a cell" means that a derived iRNA agent possesses an inhibitory activity that is at least 20% of the inhibitory activity of the iRNA agent from which it was derived in Tables 1A-1H. For example, iRNA reagents derived from the iRNA reagents of Tables 1A-1H that are capable of reducing the amount of influenza virus mRNA present in cultured influenza virus-infected human cells by 70% can be used alone in cultured influenza virus-infected human cells. The amount of influenza virus mRNA present in the human cells was reduced by 50%, so that the ability to inhibit influenza virus replication in cultured influenza virus-infected human cells is considered to be substantially preserved. Optionally, the iRNA agents of the invention can reduce the amount of influenza virus mRNA present in cultured influenza virus-infected human cells by at least 50%.

As used herein, "subject" refers to a mammalian living being who is being treated for a disease mediated by influenza virus infection. The subject can be any mammal, such as a cow, horse, mouse, rat, dog, pig, goat or primate. In preferred embodiments, the subject is a human.

Characteristics of Influenza Viruses

Influenza viruses are enveloped, negative-strand RNA viruses of the family Orthomyxoviridae. They are divided into influenza A, B and C, with influenza A being the most pathogenic and thought to be the only type capable of rearrangement with animal strains. Influenza A, B, and C can be distinguished by differences in the nucleoprotein and matrix protein. As discussed further below, influenza A subtypes are defined by mutations in their hemagglutinin (HA) or ceramidase (NA) genes, and are often distinguished by antibodies that bind to the corresponding proteins.

The influenza A virus genome consists of 10 genes distributed in 8 RNA segments. The gene encodes 10 proteins: envelope glycoproteins hemagglutinin (HA) and ceramidase (NA); matrix protein (referred to as M1 or MP in this paper); nucleoprotein (NP); three polymerases (PB1, PB2 and PA), which are constituents of RNA-dependent RNA transcriptase, also referred to herein as polymerase or polymerase complex; ion channel protein (M2), and nonstructural proteins (NS1 and NS2). For a further description of influenza A viruses and their molecular pathogenesis see Julkunen, I., et al., Cytokine and GrowthFactor Reviews, 12: 171-180, 2001. See also Fields, B., et al., Fields' Virology, 4.sup.th.ed., Philadelphia: Lippincott Williams and Wilkins; ISBN: 0781718325, 2001. The composition of the influenza B virus genome is very similar to that of influenza A, but the influenza C virus genome contains 7 RNA segments and lacks the NA gene.

The classification of influenza A viruses is based on the hemagglutinin (H1-H15) and ceramidase (N1-N9) genes. The World Health Organization (WHO) nomenclature defines each virus strain by its animal host of origin (unless specified, unless human), geographic origin, strain number, year of isolation, and HA and NA antigenic descriptions. For example, A/Puerto Rico/8/34 (H1N1) represents the A strain, isolate 8, which occurred in humans in Puerto Rico in 1934 and has HA and NA antigenic subtypes. Another example, A/Chicken/Hong Kong/258/97 (H5N1), represents the A strain, isolate 258, which occurred in chickens in Hong Kong in 1997 and has the antigenic subtype 5 of HA and antigenic subtype 1 of NA. Viruses containing HA types H1, H2, and H3 and NA types N1 and N2 have caused human epidemics.

As mentioned above, in influenza A viruses, genetic variation occurs through two main mechanisms. Antigenic drift occurs via point mutations, which often occur at positions of antigenic significance due to selective pressure from the host immune response, and antigenic shifts (also known as rearrangements), involving entire segments of the viral genome of a subtype replaced by another. Many different types of animal species, including humans, pigs, birds, horses, aquatic mammals, and others, can be infected by influenza A viruses. Some influenza A viruses are restricted to specific species and generally do not infect different species. However, some influenza A viruses can infect several different animal species, mainly birds (especially migratory waterfowl), pigs and humans. This ability is thought to be responsible for most antigenic shifts in influenza A viruses. For example, suppose a pig is infected with an influenza A virus from a human and at the same time infected with another influenza A virus from a duck. When the two different viruses were multiplied in pig cells, the genes from the human strain and those from the duck strain could "mix," resulting in a new virus with a unique combination of RNA segments. This process is called gene rearrangement (note: this type of gene rearrangement is different from the exchange of genetic information that occurs between chromosomes during meiosis).

Like other viruses and some bacterial species, influenza viruses replicate intracellularly. Influenza A viruses replicate in the epithelial cells of the upper respiratory tract. However, monocytes/macrophages and other leukocytes can also be infected. Many other cell types with cell surface glycoproteins containing sialic acid are susceptible to infection in vitro because viruses use these molecules as receptors.

Design and selection of iRNA reagents

As used herein, "disease associated with influenza virus expression" refers to any biological or pathological condition that is (1) mediated at least in part by the presence of influenza virus and (2) that a reduction in the level of influenza virus present can affect their ending. Specific diseases associated with influenza virus expression are mentioned below.

The present invention is based on the design, synthesis and production of iRNA agents capable of targeting viral genes of influenza virus, and also on the demonstration of silencing of viral genes in cultured cells in vitro after incubation with iRNA agents, and the obtained response to viral infection protective effect.

Based on sequence information and desired characteristics, iRNA agents can be rationally designed. For example, iRNAs can be designed based on the relative melting temperatures of candidate duplexes. Typically, the 5' end of the antisense strand of the duplex has a lower melting temperature than the 3' end of the antisense strand.

The invention provides compositions comprising siRNA(s) and/or shRNA(s) targeting one or more influenza virus transcripts. Because the above description of the influenza virus replication cycle demonstrates that various types of viral RNA transcripts (primary and secondary vRNA, primary and secondary viral mRNA, and viral cRNA) are present in cells infected with influenza virus, and Play an important role in the life cycle of viruses. Any of these transcripts are suitable targets for siRNA-mediated repression through direct and indirect mechanisms according to the invention. siRNAs and shRNAs targeting any viral mRNA transcript specifically reduce the level of the transcript itself in a direct manner by degrading the transcript. In addition, as discussed below, siRNAs and shRNAs targeting certain viral transcripts (eg, MP, PA, PB1) can indirectly reduce the levels of their non-specifically targeted viral transcripts. Where alternative splicing is possible, either unspliced or spliced transcripts can serve as target transcripts for MP and M2-encoding mRNAs as well as NS1 and NS2-encoding mRNAs.

Potential transcripts that can serve as targets for RNAi-based therapies according to the invention include, for example, 1) any influenza virus genome segment; 2) transcripts encoding any viral protein, including those encoding proteins PB1, PB2, PA , NP, NS1, NS2, MP, M2, HA or NA transcripts. It is understood that a single siRNA or shRNA can target transcripts in the form of vRNA, cRNA and/or mRNA. However, as suggested by Ge et al., WO 04/028471, viral mRNA may be the sole or primary target of RNAi.

Designing siRNA or shRNA for use in accordance with the present invention preferably employs certain guidelines for any particular gene target being selected. In general, it is expected that the target sequence will be specific to the virus (as compared to the host) and, preferably, be important or essential to the function of the virus. Although some viral genes, particularly those encoding HA and NA, are characterized by high mutation rates and tolerance to mutation, certain regions and/or sequences tend to be conserved. Such sequences are particularly suitable targets according to certain embodiments of the invention. As described further below, such conserved regions can be identified by consulting the literature and/or comparing influenza gene sequences, many of which are publicly available. Moreover, in many cases, the agents to be delivered to the cells of the invention may be subjected to one or more processing steps (see further discussion below) before becoming active inhibitors; in such cases, those of ordinary skill in the art It will be understood by the person that the relevant reagents are preferably designed to include the sequences necessary for their processing steps. One aspect of the present invention is the recognition that when multiple strains, subtypes, etc. (collectively referred to as variants) of infectious agents exist, with variations in their genome sequence, it is often desirable to select and/or design variants that target different variants. siRNAs and shRNAs from highly conserved regions of the body. In particular, by comparing a sufficient number of sequences and selecting highly conserved regions, it is possible to target multiple variants with a single siRNA, the duplex portion of which includes the highly conserved regions. Typically, the region should be of sufficient length to include the entire duplex portion of the siRNA (e.g., 19 nucleotides), and optionally, one or more 3' overhangs, although longer than the entire duplex portion. Shorter and longer regions may also be used (eg, 15, 16, 17 or 18 nucleotides). According to some embodiments of the invention, a region is highly conserved among variants if it is identical across variants. According to certain embodiments of the invention, if a region (whether or not the length is included within the duplex portion of the siRNA, e.g. 15, 16, 17, 18 or preferably 19 nucleotides) has at most 1 core between variants Nucleotides (ie, 0 or 1 nucleotide) differ if they are highly conserved. According to certain embodiments of the invention, a region is highly conserved if it differs by at most 2 nucleotides (ie 0, 1 or 2 nucleotides) between variants. According to certain embodiments of the invention, a region is highly conserved if it differs between variants by up to 3 nucleotides (ie 0, 1, 2 or 3 nucleotides). According to some embodiments of the invention, the siRNA comprises a duplex portion targeting a region in at least 5 variants, at least 10 variants, at least 15 variants, at least 20 variants, at least 25 variants, at least 30 variants, at least 40 variants, or at least 50 variants or more are highly conserved.

To determine whether a region is highly conserved among variants, the following method can be used. A member of the set of sequences is selected as the base sequence, that is, the sequence against which the other sequences are compared. Typically, the length of the base sequence is the desired length of the duplex portion of the siRNA, eg 15, 16, 17, 18 or preferably 19 nucleotides. According to various embodiments of the invention, the base sequence may be either a sequence in the set being compared, or a derived consensus sequence, for example derived by determining the most common nucleotide at each position in a set of sequences consensus sequence.

After the base sequence is selected, each member sequence of the set of multiple variants is compared to the base sequence. The number of differences in that sequence region between the base sequence and any member of the set of multiple variants can be used to determine whether the base sequence and the member are highly conserved over a particular region of interest. As mentioned above, in various embodiments of the invention, if the number of sequence differences between two regions is 0; 0 or 1, 0, 1 or 2; or 0, 1, 2 or 3, then This region is considered to be highly conserved. Where differences occur, the siRNA sequence can be selected to be identical to the base sequence or one of the other sequences. Typically, nucleotides that are present in the base sequence are selected. However, in certain embodiments of the invention, in particular if the nucleotide present at a particular position in the second sequence of the comparison set is found in more of the compared sequences than the nucleotide in the base sequence, then the Produce the same siRNA as the second sequence. Additionally, according to certain embodiments of the invention, the consensus nucleotide (the most prevalent nucleotide) at which the difference occurs can be used if it differs from the nucleotide found in the base sequence . Note that this may make the obtained sequence different from all the sequences being compared (as can a consensus sequence be used as a base sequence).

The inventors found that most of the sequences selected using the design parameters described below were effective in inhibiting viral replication after introduction of siRNA or shRNA and were tested as described below.

Based on the results shown herein, the present invention provides iRNA agents that reduce influenza virus replication in cultured influenza virus-infected cells as well as in subjects such as mammals, eg, humans. Tables 1A-1H provide exemplary iRNA agents targeting influenza virus. Tables 1A, C, D and E list siRNAs containing no nucleotide modifications except for the phosphorothioate bond between the 3' end and the penultimate thymidine. Tables 1B and H list siRNAs in which all nucleotides containing pyrimidine bases are 2'-O-methyl-modified nucleotides in the sense strand, and all nucleotides in the 5'-ua-3' sequence background Uridine as well as cytidine in the context of all 5'-ca-3' sequences are 2'-O-methyl-modified nucleotides in the antisense strand, except for AL-DP-2295 which has duplex determination, Except for the iRNAs of AL-DP-2301 and AL-DP-2302, all of the uridines in the context of the 5'-ug-3' sequence were 2'-O-methyl-modified nucleotides. These latter siRNAs did not exhibit the sequence motif 5'-ua-3' or 5'-ca-3', and after incubation of these reagents in mouse serum, analysis of these degraded fragments showed that the sequence motif 5' -ug-3' is the main site of endonuclease attack.

Based on these results, the present aspects specifically provide iRNA agents comprising a sense strand and an antisense strand, the sense strand having at least 15 contiguous nucleotides of the sense strand sequence of the agents shown in Tables 1A-1H, The antisense strand has at least 15 contiguous nucleotides of the antisense strand of the reagents shown in Tables 1A-1H.

The iRNA reagents shown in Tables 1A-1H consist of two strands 19 nucleotides in length that are complementary or identical to the target sequence, plus a 3'- TT overhang. The invention provides reagents comprising at least 15 or at least 16, 17 or 18 or 19 contiguous nucleotides of these sequences. However, while these lengths may potentially be optimal, it is not meant to limit iRNAs to these lengths. Those of skill in the art will appreciate that shorter or longer iRNA agents are equally effective because, within a certain length, efficacy is a function of the nucleotide sequence rather than the length of the sequence. For example, Yang, et al., PNAS 99 :9942-9947 (2002) demonstrated similar efficacy for iRNA agents between 21 and 30 base pairs in length. Others have also demonstrated that iRNA reagents as small as about 15 base pairs in length can effectively silence genes (Byrom, et al., "Inducing RNAi with siRNA Cocktails generated by RNase III" Tech Notes 10(1), Ambion, Inc., Austin, TX).

Therefore, the present invention can and contemplates selecting a part of the sequence between 15-19 nucleotides from the sequences provided in Tables 1A-1H for obtaining a sequence derived from one of the sequences provided in Tables 1A-1H iRNA reagents. Alternatively, one or several nucleotides may be added to a sequence provided in Tables 1A-1H, or to a reagent containing 15 contiguous nucleotides from one of these reagents, preferably , but not necessarily, in such a way that the added nucleotides are complementary to the respective sequences of the target gene, such as an influenza virus gene. For example, combining the first 15 nucleotides of a reagent with the 8 nucleotides found at the 5' end of the sequence in influenza virus mRNA results in a reagent with 23 nucleotides in the sense and antisense strands. All such derivatized iRNA agents are included in the iRNA agents of the present invention, so long as they substantially retain the ability to inhibit influenza virus replication in cultured influenza virus-infected human cells.

The antisense strand length of the iRNA agent should be equal to or at least 14, 15, 16, 17, 18, 19, 25, 29, 40 or 50 nucleotides in length. It should be equal to or less than 60, 50, 40 or 30 nucleotides in length. Preferred length ranges are 15-30, 17-25, 19-23 and 19-21 nucleotides.

The sense strand length of the iRNA agent should be equal to or at least 14, 15, 16, 17, 18, 19, 25, 29, 40 or 50 nucleotides in length. It should be equal to or less than 60, 50, 40 or 30 nucleotides in length. Preferred length ranges are 15-30, 17-25, 19-23 and 19-21 nucleotides.

The double-stranded portion of the iRNA agent should be equal to or at least 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 29, 40, or 50 nucleotide pairs in length. It should be equal to or less than 60, 50, 40 or 30 nucleotide pairs in length. Preferred length ranges are 15-30, 17-25, 19-23 and 19-21 nucleotide pairs.

Typically, the iRNA agents of the invention include a region that is sufficiently complementary to the corresponding influenza virus gene and are sufficiently long in nucleotides that the iRNA agent or fragment thereof can mediate down-regulation of the influenza virus gene. Perfect complementarity between the iRNA agent and the target gene is not required, but the identity must be sufficient so that the iRNA agent or its cleavage products can direct sequence-specific silencing, such as cleavage of influenza virus RNA by RNAi.

Accordingly, iRNA agents of the invention include agents comprising a sense strand and an antisense strand each comprising a sequence of at least 16, 17, or 18 nucleotides, as described below defined, substantially identical to a sequence in Tables 1A-1H, except that no more than 1, 2 or 3 nucleotides of each strand, respectively, are substituted by other nucleotides (e.g. uracil in place of adenosine), Moreover, the ability to inhibit influenza virus replication in cultured influenza virus-infected human cells is substantially retained, respectively. Thus, these reagents have at least 15 nucleotides identity to a sequence in Tables 1A-1H, but introduce 1, 2 or 3 base errors relative to the target influenza RNA sequence or between the sense and antisense strands. match. Mismatches to the target influenza virus RNA sequence, especially in the antisense strand, are most tolerated in the terminal region and, if present, are preferably within 1 or more regions of the terminal, e.g., at 5 Within 6, 5, 4 or 3 nucleotides of the ' and/or 3' end, most preferably within 6, 5, 4 or 3 nucleotides of the 5' end of the sense strand or the 3' end of the antisense strand . The sense strand only needs to be sufficiently complementary to the antisense strand to maintain the fully double-stranded character of the molecule.

Preferably, the sense and antisense strands are selected so that the iRNA agent includes single-stranded or unpaired regions at one or both ends of the molecule. Thus, an iRNA agent contains a sense strand and an antisense strand, preferably paired to include an overhang, such as one or both 5' or 3' overhangs, but preferably a 3' overhang of 2-3 nucleotides. Most embodiments have a 3' overhang. Preferred siRNA agents have single-stranded overhangs, preferably 3' overhangs, of 1-4 or preferably 2 or 3 nucleotides in length at one or both ends of the iRNA agent. Overhangs can be the result of one strand being longer than the other, or two strands of the same length being interleaved. The unpaired nucleotides forming the overhang may be ribonucleotides or deoxyribonucleotides, preferably thymidine. The 5' end is preferably phosphorylated, or may not be phosphorylated.

The preferred length of the duplexed region is 15 to 30 nucleotides, most preferably 18, 19, 20, 21, 22 and 23 nucleotides, eg within the range of siRNA agents as discussed above. SiRNA agents can be similar in length and structure to natural Dicer-processed products from long dsRNAs. In one embodiment, the two strands of the siRNA agent are linked, such as covalently linked. Hairpin structures or other single-stranded structures providing the desired double-stranded region, preferably a 3' overhang, are also within the scope of the invention.

Evaluation of Candidate iRNA Reagents

As noted above, the present invention provides a system for identifying siRNAs useful as inhibitors of influenza virus infection and/or replication. As mentioned above, because shRNAs are processed in cells to generate duplex portions of siRNAs that have the same sequence as the stem structure of shRNAs, the system can also be used to determine the inhibitory effect of influenza virus infection. agent shRNAs. For descriptive purposes, this section refers to siRNAs, but the system also includes corresponding shRNAs. Specifically, the present invention demonstrates the successful preparation of siRNAs targeting viral genes to suppress or inhibit viral infection and/or replication. The techniques and reagents described herein can be conveniently used to design potential new siRNAs targeting other genes or gene regions, and to readily test their activity in inhibiting influenza virus infection and/or replication as described above. Influenza viruses are expected to continue to mutate and rearrange, so it is expected that new and differently targeted siRNAs will continue to be developed and tested.

In various embodiments of the invention, prior to, simultaneously with, or after transfection (e.g., within minutes, hours, or up to several days) with the influenza virus genome or a portion thereof, or after transfection with the influenza virus genome By introducing candidate siRNA(s) into cells (such as by exogenous administration (exogeneous adminstration) or by introducing a vector or construct directing the exogenous synthesis of siRNA into cells) or into a test animal prior to, concurrently with, or after infection, or into a test animal, to test potential influenza virus inhibitors. Alternatively, influenza virus inhibitors can be tested by introducing candidate siRNA(s) into cells or experimental animals that are productively infected with influenza virus (i.e., cells capable of producing progeny viruses) . Subsequently, the ability of the candidate siRNA(s) to reduce the level of the target transcript and/or suppress or suppress one or more aspects of the viral life cycle, such as viral replication, pathogenicity and/or infectivity, is assessed. For example, viral particle production and/or viral protein production can be assessed directly or indirectly using methods well known in the art.

Cells or test animals that have been delivered with the siRNA composition of the present invention (test cells/animals) can be compared to similar or comparable cells or test animals that have not received the composition of the present invention (control cells/animals, e.g., do not receive siRNA or receive Control siRNA such as cells/animals targeting non-viral transcripts such as GFP) for comparison. The susceptibility of test cells/animals to influenza virus infection can be compared to the susceptibility of control cells/animals to infection. The production of viral proteins and/or progeny virus can be compared in test cells/animals relative to control cells/animals. Other indicators (indicia) such as virus infectivity, replication, and pathogenicity can also be compared. In vitro antiviral detection standards can be used to inhibit viral plaques, viral cytopathic effects (cytopathic effect, CPF), viral hemagglutinin or other proteins, and inhibition of viral production. CPE can be measured visually as well as by dye uptake. See eg Sidwell, R.W. and Smee, D.F, "In vitro and in vivo assay systems for study of influenza virus inhibitors" Antiviral Res 2000, 48:1. Typically, the test cell/animal and the control cell/animal are from the same species and, for the cells, may be of a similar or identical cell type. For example, cells from the same cell line can be compared. When the test cells are primary cells, generally, the control cells should also be primary cells. Usually the same strain of influenza virus is used to compare test cells/animals and control cells/animals.

For example, the ability of a candidate siRNA to inhibit influenza virus production can be conveniently determined by: (i) delivering the candidate siRNA to cells (before, simultaneously with, or after exposure to influenza virus); (ii) assessing viral hemagglutinin production; and (iii) comparing the amount of hemagglutinin produced in the presence of siRNA to the amount of hemagglutinin produced in the absence of siRNA. (The test need not include controls lacking siRNA, but can utilize previous information on the amount of hemagglutinin produced in the absence of inhibition). Significantly lower amounts of hemagglutinin indicate reduced virus production. This assay can be used to test siRNAs targeting any viral transcript and is not limited to siRNAs targeting transcripts encoding viral hemagglutinin.

The ability of a candidate siRNA to reduce the level of a target transcript can also be measured by measuring the amount of the target transcript using, for example, Northern blotting, nuclease protection assays, reverse transcription (RT)-PCR, real-time RT-PCR, microarray analysis, etc. to evaluate. The ability of a candidate siRNA to inhibit production of a polypeptide encoded by a target transcript (at the transcriptional or post-transcriptional level) can be measured using a variety of antibody-based methods including, but not limited to, Western blots, immunoassays, ELISAs, Flow cytometry, protein microarray, etc. In general, any method that measures the amount of a target transcript or a polypeptide encoded by a target transcript can be used.

In general, certain preferred influenza virus inhibitors reduce the level of a target transcript by at least about 2-fold relative to the level that would be present in the absence of the inhibitor (e.g., in the same cells lacking the inhibitor), preferably At least about 4 times, more preferably at least about 8 times, at least about 16 times, at least about 64 times or even higher degrees. In general, certain preferred influenza virus inhibitors inhibit viral replication such that the level of replication is at least about 2-fold, preferably at least about 4-fold lower in cells containing the inhibitor than in control cells not containing the inhibitor, More preferably at least about 8-fold, at least about 16-fold, at least about 64-fold, at least about 100-fold, at least about 200-fold, or to an even higher degree.

Certain preferred influenza virus inhibitors inhibit viral replication, thereby preventing the development of detectable viral titers for at least 24 hours, at least 36 hours, at least 48 hours, or at least 60 hours after administration of the siRNA and infection of the cells. Certain preferred influenza virus inhibitors prevent (ie, reduce to undetectable levels) or significantly reduce viral replication for at least 24 hours, at least 36 hours, at least 48 hours, or at least 60 hours after administration of the siRNA. According to various embodiments of the present aspect, the significant reduction in viral replication is a reduction to less than about 90% of the level that might occur in the absence of siRNA, a reduction to less than about 75% of the level that might occur in the absence of siRNA, a reduction to no Less than about 50% of the level that might be present in the absence of siRNA, reduced to less than about 25% of the level that might be present in the absence of siRNA, or reduced to less than about 10% of the level that might be present in the absence of siRNA. Reduction in viral replication can be measured using any suitable method including, but not limited to, measuring HA titers.

iRNA Reagent Stability Testing, Modification, and Retesting

Candidate iRNA agents can be assessed for stability, eg, susceptibility to endonuclease or exonuclease cleavage, such as when the iRNA agent is introduced into a subject. Methods can be used to identify sites susceptible to modification, particularly cleavage, eg, by components found in the subject. The methods include isolating and identifying a candidate iRNA agent following incubation with an isolated biological medium such as serum, plasma, saliva, cerebrospinal fluid, or cell or tissue homogenate in vitro, or contacting a subject with a candidate iRNA agent in vivo, The most abundant fragments formed by the degradation of candidate iRNA reagents identify the sites of ease of cleavage. Such methods are found, for example, without limitation, in co-owned International Application No. PCT/US2005/018931, filed May 27, 2005.

Once cleavage-susceptible sites are identified, further iRNA agents can be designed and/or synthesized, wherein the potential cleavage site is made resistant to cleavage, for example by introducing a 2' modification at the cleavage site, such as 2'-O- methyl. The stability of such further iRNA agents can be retested, and the process can be repeated until the iRNA agent is found to exhibit the desired stability.

in vivo test

The in vivo functionality of iRNA agents determined to inhibit influenza virus gene expression can be tested in animal models (eg, in mammals, eg, in mice or rats). For example, an iRNA agent can be administered to an animal and evaluated for its biodistribution, stability, and its ability to inhibit influenza virus replication or reduce a biological or pathological process mediated at least in part by influenza virus.

The iRNA agent can be administered directly to the target tissue, such as by injection, or the iRNA agent can be administered to animal models in the same manner as it is administered to humans. Preferably, the iRNA agent is delivered to the subject's airway, such as intranasally.

The intracellular distribution of iRNA can also be assessed. The evaluation includes determining whether the iRNA agent is taken up into the cell. The evaluation also includes determining the stability (eg, half-life) of the iRNA. By using iRNA reagents linked to traceable labels (e.g., fluorescent labels such as fluorescein; radioactive labels such as ³⁵ S, ³² P, ³³ P, or ³ H; gold particles; or antigen particles for immunohistochemistry) , can facilitate the evaluation of iRNA agents in vivo.

The ability of iRNA agents to downregulate influenza virus replication can be assessed. The level of expression of influenza virus genes in vivo can be measured, for example, by in situ hybridization, or by isolating RNA from tissue either before or after exposure to an iRNA agent. In cases where animal sacrifice is required to obtain tissue, untreated control animals can be used as controls. Influenza virus RNA can be detected by any desired method including, but not limited to, PR-PCR, Northern blot, branched DNA analysis, or ribonuclease protection assay. Alternatively, or in addition, influenza virus gene expression can be monitored by Western blot analysis or plaque formation assays on tissue extracts treated with iRNA reagents.

Potential inhibitors of influenza virus can be tested using any of the various animal models that have been established. Compositions comprising candidate siRNA(s) capable of directing the siRNA in a host cell or in a cell engineered or manipulated to contain the candidate siRNA are administered to the animal prior to, concurrently with, or following infection with influenza virus Synthetic constructs or vectors. The compositions are evaluated for their ability to prevent viral infection and/or delay or prevent the onset of influenza-associated symptoms and/or lessen the severity of influenza virus-infected animals not receiving potential influenza virus inhibitors. Such models include, but are not limited to, murine (murine), chick, ferret (ferret), and non-human primate models for influenza infection, which are known in the art and can be used to test potential Efficacy of influenza therapies and vaccines. See eg Sidwell, R.W. and Smee, D.F, supra referenced. The model can use naturally occurring strains of influenza virus and/or strains that have been modified or adapted to survive in a particular host (eg, WSN or PR8 strains that are adapted to replicate in mice). The above animal models can also be used to determine the concentrations required to achieve a certain desired effect (eg, EC50).

iRNA chemistry

Isolated iRNA agents, eg, dsRNA agents that mediate RNAi to inhibit influenza virus gene expression are described herein.

The RNA agents discussed here include RNA that is otherwise unmodified and RNA that has been modified to improve its efficacy, and also includes polymers of nucleoside surrogates. Unmodified RNA refers to a molecule in which the components of nucleic acid, ie sugar, base and phosphate moieties, are identical or substantially identical to those naturally occurring, preferably in the human body. Rare or uncommon but naturally occurring RNAs are referred to as modified RNAs in the prior art, see, for example, Limbach et al., Nucleic Acids Res. 22 :2183-2196, 1994. Such rare or uncommon RNAs, commonly referred to as modified RNAs (as if since they are often the product of post-transcriptional modifications), are within the scope of the term "unmodified RNA" herein. The modified RNA described herein refers to a molecule whose components of nucleic acid, ie, sugar, base and phosphate moiety, are different from naturally occurring components, preferably from those naturally occurring in the human body. Although they are referred to as modified "RNA", molecules that are not RNA can of course be included due to the modification. Nucleoside surrogates are molecules in which the ribose phosphate backbone is replaced by a non-ribose phosphate construct that allows the bases to exist in the correct spatial relationship, thereby allowing hybridization and analogues with ribose phosphate backbones (e.g., uncharged ribose phosphate backbones) species) are basically similar. This article will discuss the above examples.

The modifications described herein can be introduced into any of the double-stranded RNAs described herein, as well as RNA-like molecules such as iRNA agents. It may be preferred to modify one or both of the antisense and sense strands of the iRNA agent. Because nucleic acids are polymers of subunits or monomers, many of the modifications described below occur at recurring positions within nucleic acids, such as modifications to bases or phosphate sites or non-linking O's of phosphate moieties. In some cases, the modification occurs at all subject positions of the nucleic acid, but in many (indeed most) cases this is not the case. For example, the modification may only occur at the 3' or 5' terminal position, may only occur at the terminal region, for example at the position of the terminal nucleotide or within the last 2, 3, 4, 5 or 10 nucleotides of the strand. Occurs in the double-stranded region, occurs in the single-stranded region, or in both regions. For example, a phosphorothioate modification at a non-linking O position may occur only at one or both termini, may only occur at a terminal region, for example at the position of a terminal nucleotide or the last 2, 3, 4, 5 or Within 10 nucleotides, or can occur only in single- and double-stranded regions, especially at the ends. Likewise, modifications can occur on the sense strand, the antisense strand, or both strands. In some cases, the sense and antisense strands have the same modification or have the same type of modification, but in other cases, the sense and antisense strands have different modifications, for example, in some cases only the One strand is modified, for example, the sense strand only.

The two main purposes of introducing modifications into iRNA agents are to have degradation stability in biological environments and to improve pharmacological properties such as pharmacodynamic properties, which are discussed further below. Other suitable modifications to the sugar, base or backbone of iRNA agents are described in co-owned PCT Application No. PCT/US2004/01193, filed January 16, 2004. iRNA agents can include non-naturally occurring bases, such as those described in co-owned PCT Application No. PCT/US2004/011822, filed April 16, 2004. iRNA agents can include non-naturally occurring sugars, such as non-carbohydrate cyclic carrier molecules. Exemplary properties of non-naturally occurring sugars for use in iRNA agents are described in co-owned PCT Application No. PCT/US2004/11829, filed April 16, 2003.

iRNA agents may contain internucleotide linkages (eg, chiral phosphorothioate linkages) for increased nuclease resistance. Additionally, or alternatively, the iRNA agent may comprise a ribose mimetic to increase nuclease resistance. Exemplary internucleotide linkages and ribose mimetics for increasing nuclease resistance are described in co-owned PCT Application No. PCT/US2004/07070, filed March 8, 2004.

iRNA agents can include ligand-binding monomer subunits and monomers suitable for oligonucleotide synthesis. Exemplary monomers are described in co-owned US Application No. 10/916,185, filed August 10, 2004.

iRNA agents can have a ZXY structure, such as described in co-owned PCT Application No. PCT/US2004/07070, filed March 8, 2004.

iRNA agents can be complexed with amphoteric components. Exemplary amphoteric components for use in iRNA agents are described in co-owned PCT Application No. PCT/US2004/07070, filed March 8, 2004.

In another embodiment, an iRNA agent can be complexed with a delivery agent that has the properties of a modular complex. The complex includes a carrier agent linked to one or more (preferably two or more, more preferably three) of the following agents: (a) a condensing agent (e.g., capable of attracting Such as reagents that bind nucleic acids; (b) fusogenic agents (e.g., agents capable of fusion and/or transport across cell membranes; and (c) targeting groups, such as cell or tissue targeting agents such as lectins, glycoproteins, Lipids or proteins, such as antibodies that bind to specific cell types. iRNA agents complexed with delivery agents are described in co-owned PCT Application No. PCT/US2004/07070, filed March 8, 2004.

iRNA agents may contain non-canonical pairings, such as between the sense and antisense sequences of an iRNA duplex. Exemplary features of non-canonical iRNA agents are described in co-owned PCT Application No. PCT/US2004/07070, filed March 8, 2004.

Enhanced nuclease resistance

iRNA agents, such as iRNA agents targeting influenza virus, may have enhanced nuclease resistance.

One approach to increasing resistance, as described in co-owned US Application No. 60/559,917, filed May 4, 2004, is to identify the cleavage site and modify the site to inhibit cleavage. For example, dinucleotides 5'-ua-3', 5'-ca-3', 5'-ug-3', 5'-uu-3' or 5'-cc-3' can serve as cleavage sites . In certain embodiments, all pyrimidines in the iRNA agent carry 2'-modifications in the sense strand, the antisense strand, or both, such that the iRNA agent has increased resistance to endonucleases. Nuclease resistance can also be enhanced by modifying the 5' nucleotide such that, for example, urea in at least one 5'-uridine-adenine-3' (5'-ua-3') dinucleotide glycoside is a 2'-modified nucleotide; at least one 5'-cytidine in a 5'-cytidine-adenine-3'(5'-ca-3') dinucleotide is a 2'-modified nucleoside acid; at least one 5'-uridine-guanine-3' (5'-ug-3') dinucleotide in which the 5'-uridine is a 2' modified nucleotide; at least one 5'-uridine The 5'-uridine in the -uridine-3'(5'-uu-3') dinucleotide is a 2'-modified nucleotide; or at least one 5'-cytidine-cytidine-3'( The 5'-cytidine in the 5'-cc-3') dinucleotide is a 2'-modified nucleotide, as described in co-applicants' International Application PCT/US2005/018931 filed May 27, 2005 stated. An iRNA agent can include at least 2, at least 3, at least 4, or at least 5 such dinucleotides. In a particularly preferred embodiment, the 5' nucleotides in all occurrences of the 5'-ua-3' and 5'-ca-3' sequence motifs in the sense strand, the antisense strand, or both strands are modified of nucleotides. Preferably the 5' nucleotides in all occurrences of the 5'-ua-3', 5'-ca-3' and 5'-ug-3' sequence motifs in the sense strand, the antisense strand or both strands are Modified nucleotides. More preferably all pyrimidine nucleotides of the sense strand are modified nucleotides and all occurrences of 5' nucleotides in the 5'-ua-3' and 5'-ca-3' sequence motifs of the antisense strand The antisense strand lacks the 5'-ua-3' and 5'-ca-3' motifs for modified nucleotides, or in all cases where there is a 5'-ug-3' sequence.

Preferably, 2'-modified nucleotides include, for example, 2'-modified ribose units. For example, the 2'-hydroxyl group (OH) can be modified or replaced by a variety of different "oxy" or "deoxy" substituents.

Examples of "oxy"-2' hydroxy group modifications include alkoxy or aryloxy (OR, such as R=H, alkyl, cycloalkyl, aryl, aralkyl, heteroaryl, or sugar); poly Ethylene glycol (PEG); O(CH ₂ CH ₂ O) _n CH ₂ CH ₂ OR; "locked" nucleic acids (LNA), in which the 2' hydroxyl group is linked, for example, by a methylene bridge, to the 4' carbon of the same ribose sugar; O-amine (O-AMINE) and aminoalkoxy, O(CH ₂ ) _n amine, (for example, amine=NH ₂ , alkylamino, dialkylamino, heterocyclylamino, arylamino, diarylamino , heteroarylamino, or diheteroarylamino, hexamethylenediamine, polyamine). Notably, compared to oligonucleotides modified with strong phosphorothioates, oligonucleotides containing only methoxyethyl groups (MOE), (OCH ₂ CH ₂ OCH ₃ , a PEG derivative), Nucleotides exhibit nuclease stability.

"Deoxy" modifications include hydrogen (i.e., deoxyribose, which is of particular relevance to the protruding portion of some dsRNAs); halo (e.g., fluorine); ammonia (e.g., _NH2 , alkylamino, dialkylamino, heterocyclyl , arylamino, diarylamino, heteroarylamino, diheteroarylamino, or amino acid); NH(CH ₂ CH ₂ NH) _n CH ₂ CH ₂ -amine (amine=NH ₂ , alkylamino, di Alkylamino, heterocyclylamino, arylamino, diarylamino, heteroarylamino or diheteroarylamino), -NHC(O)R (R = alkyl, cycloalkyl, aryl, aralkyl group, heteroaryl or sugar), cyano, mercapto; alkyl-thio-alkyl; thioalkoxy; and alkyl, cycloalkyl, aryl, alkenyl and alkynyl, which can be replaced by, for example, amino Functional groups are optionally substituted.

Preferred substituents are 2'-methoxyethyl, 2'- _OCH3 , 2'-O-allyl, 2'-C-allyl and 2'-fluoro.

The inclusion of furanose in the oligonucleotide backbone also reduces endonuclease cleavage. iRNA agents can be further modified by introducing 3' cationic groups, or by converting nucleosides at the 3'-terminus using 3'-3' linkages. In a further alternative, the 3'-terminus can be blocked with an aminoalkyl group such as 3'C5-aminoalkyldT. Other 3' conjugates can also inhibit 3'-5' exonuclease cleavage. Without being bound by any theory, 3' conjugates such as naproxen or ibuprofen inhibit exonuclease cleavage by sterically hindering the binding of exonucleases to the 3'-terminus of oligonucleotides . Even small alkyl chains, aryl groups, or heterocyclic conjugates or modified sugars (D-ribose, deoxyribose, glucose, etc.) can block 3'-5'-exonucleases.

Nucleolytic cleavage can be inhibited by introducing a phosphate-linker modification, such as a phosphothiol ester linkage. Therefore, preferred iRNA agents include nucleotide dimers that are enriched or all in a specific chiral form of a modified phosphate group that is normally occupied by an oxygen contain heteroatoms at non-bridge positions. The heteroatom may be S, Se, _Nr2 or _Br3 . When the heteroatom is S, it is preferably enriched or entirely chiral (chirally pure) Sp linkages. Enriched means at least 70, 80, 90, 95 or 99% of the preferred form. Modified phosphate linkers are particularly effective at inhibiting exonuclease cleavage when introduced near the 5'- or 3'-terminal position of the iRNA agent, especially at the 5'-terminal position.

The 5' conjugate also inhibits 5'-3' exonuclease cleavage. Without being bound by any theory, 5' conjugates such as naproxen or ibuprofen inhibit exonuclease activity by sterically hindering the binding of exonucleases to the 5'-terminus of oligonucleotides. crack. Even small alkyl chains, aryl groups, or heterocyclic conjugates or modified sugars (D-ribose, deoxyribose, glucose, etc.) can block 3'-5'-exonucleases.

When the double-stranded iRNA agent comprises a single-stranded nucleotide overhang on at least one terminus, the iRNA agent can have enhanced resistance to nucleases. In a preferred embodiment, the nucleotide overhang comprises 1-4, preferably 2-3 unpaired nucleotides. In a preferred embodiment, the unpaired nucleotides of the single-stranded overhang directly connected to the terminal nucleotide pair contain purine bases, and the terminal nucleotide pair is a G-C pair, or the last 4 complementary cores At least two of the nucleotide pairs are G-C pairs. In a further embodiment, the nucleotide overhang may have 1 or 2 unpaired nucleotides, and in an exemplary embodiment, the nucleotide overhang is 5'-gc-3'. In a preferred embodiment, the nucleotide overhang is at the 3'-end of the antisense strand. In one embodiment, the iRNA agent includes a 5'-cgc-3' motif at the 3'-end of the antisense strand, forming a 2-nt overhang 5'-gc-3'.

Thus, iRNA agents may contain modifications to inhibit degradation by nucleases such as endonucleases or exonucleases found in the subject. These monomers are referred to herein as NRMs or nuclease resistance promoting monomers, and the corresponding modifications are referred to as NRM modifications. In many cases, these modifications may also modulate other properties of the iRNA agent, such as the ability to interact with proteins such as transporters such as serum albumin or with RISC members, or the ability of the first and second sequences to form duplexes with each other or The ability to form a duplex with another sequence such as a target molecule.

One or more different NRM modifications can be introduced into the iRNA agent, or into the sequence of the iRNA agent. NRM modification can be used more than once in the sequence or in the iRNA reagent.

NRM modifications include some that can be present only at the ends and others that can be present at any position. Some NRM modifications can inhibit hybridization, so it is preferred to use them only in the terminal regions, and more preferably not to target cleavage sites or regions in the sequence of the subject sequence or gene, especially in the antisense strand. They can be used anywhere on the sense strand as long as hybridization between the two strands of the dsiRNA agent can be adequately maintained. In some embodiments, it is preferable to place the NRM at the cleavage site or region of the sense strand, thereby minimizing off-target silencing.

In most cases, NRM modifications can be in different distributions depending on whether they are contained in the sense or antisense strand. If included in the antisense strand, modifications that interfere with or inhibit endonuclease cleavage should not be inserted into regions that are subject to RISC-mediated cleavage, such as the cleavage site or region (e.g. Elbashir et al., 2001, Genes and Dev .15:188, which is incorporated herein by reference). Cleavage of the target occurs approximately in the middle of the 20 or 21 nt antisense strand, or approximately 10 or 11 nucleotides upstream of the first nucleotide of the target mRNA (which is complementary to the antisense strand). A cleavage site as used herein refers to the nucleotides on both sides of the cleavage site on the strand of the target or iRNA agent to which it hybridizes. Cleavage region refers to nucleotides within 1, 2 or 3 nucleotides in either direction of the cleavage site.

The modification may be introduced into the terminal region, for example at the terminal position of the sense or antisense strand or at 2, 3, 4 or 5 positions from the terminal.

Tethered Ligands

The properties of an iRNA agent, including its pharmacological properties, can be influenced and tailored, for example, by introducing ligands such as tethering ligands. In addition, the pharmacological properties of the iRNA agent, when the iRNA agent either already or has a ligand tethered, is improved by the introduction of the ligand.

Various constituents such as ligands can be tethered into the iRNA reagent or as formulation excipients or additives, for example, into a ligand-binding monomer subunit carrier. Examples of ligand-binding monomer subunits are described below, but these are only preferred examples, and the constituent elements can also be attached to the iRNA agent at other positions.

A preferred constituent is a ligand, which is directly or indirectly linked, preferably covalently, to the carrier via an intervening plug. In preferred embodiments, the ligand is attached to the carrier via an intervening plug. The ligand or tethered ligand may be present on the ligand-binding monomer when the ligand-binding monomer is introduced into the growing chain. In some embodiments, the ligand can be introduced into the "precursor" ligand-binding monomer subunit after the "precursor" ligand-binding monomer subunit has been introduced into the growing chain. For example, a monomer with, for example, _an amino-terminal plug, such as TAP-( _CH2 ) _nNH2 , can be inserted into the growing sense or antisense strand. In subsequent manipulations, that is, after the precursor monomer subunits have been inserted into the chain, ligands having electrophilic groups such as pentafluorophenyl ester or aldehyde groups are then passed through the electrophilic groups of the ligands The way that the group is connected to the terminal nucleophilic group of the precursor ligand-binding monomer subunit plug is connected to the precursor ligand-binding monomer.

In preferred embodiments, the ligand alters the distribution, targeting or half-life of the iRNA agent into which it is introduced. In a preferred embodiment, the ligand provides enhanced affinity for a selected target, e.g., a molecule, cell, or cell type, compared to, e.g., a species in the absence of the ligand. , a compartment such as a cell compartment or an organ compartment, tissue, organ or body part.

Preferred ligands can improve the transport, hybridization and specificity of the resulting natural or modified oligoribonucleic acid, or polymeric molecules containing any combination of monomers and/or natural or modified ribonucleotides described herein and can improve Its nuclease resistance.

Ligands typically include, for example, therapeutic modulators to facilitate absorption; diagnostic compounds or reporter groups, such as to monitor distribution; cross-linking agents; agents that confer nuclease resistance; and natural or unusual nucleic acid bases. Common examples include lipophilic molecules, lipids, lectins, steroids (e.g. arbutol, hecigenin, diosgenin), terpenes (e.g. triterpenes such as sarsaparin Lithocholic acid derived from epixetin, epixetol, lithocholic acid), vitamins, carbohydrates (such as dextran, pullulan, chitin, chitosan, inulin, cyclodextrin, or hyaluronic acid) , proteins, protein binders, integrin targeting molecules, polycations, peptides, polyamines, and peptide analogs.

The ligand may be a naturally occurring, recombinant or synthetic molecule, such as a synthetic polymer such as a synthetic polyamino acid. Examples of polyamino acids include polylysine (PLL), poly-L-aspartic acid, poly-L-glutamic acid, styrene-maleic anhydride copolymer, poly(L-propiolactone- Co-glycolized) copolymer, divinyl ether-maleic anhydride copolymer, N-(2-hydroxypropyl)methacrylamide copolymer (HMPA), polyethylene glycol (PEG), polyvinyl alcohol (PVA), polyurethane, poly(2-ethylacrylic acid), N-isopropylacrylamide polymer, or polyphosphazine. Examples of polyamines include: polyethyleneimine, polylysine (PLL), spermine, spermidine, polyamines, pseudopeptide polyamines, peptidomimetic polyamines, dendrimer polyamines, arginine Acids, amidines, protamines, cationic components such as cationic lipids, cationic porphyrins, quaternary ammonium salts of polyamines, or alpha-helical peptides.

Ligands can also include targeting groups such as thyrotropin, melanocyte-stimulating hormone, surfactant protein A, mucin sugars, glycosylated polyamino acids, transferrin, bisphosphonates, polyglutamate , polyarginine salts, or RGD peptides or RGD peptide analogs and other cell or tissue targeting agents.

Ligands can be proteins, such as glycoproteins, lipoproteins such as low-density lipoprotein (LDL), albumins such as human serum albumin (HSA), or peptides such as molecules with specific affinity for co-ligands, which interact with cancer cells, endothelial Antibodies that bind to specific cell types such as cells or bone cells. Ligands may also include hormones or hormone receptors. They may also include non-peptides such as cofactors, polyvalent lactose, polyvalent galactose, N-acetyl-galactosamine, N-acetyl-glucosamine, polyvalent mannose or polyvalent trehalose. The ligand can be, for example, lipopolysaccharide, p38 MAP kinase activator, or NF-κB activator.

The ligand can be a substance, such as a drug, that increases the amount of iRNA agent absorbed into the cell by, for example, disrupting the cytoskeleton of the cell, eg, by disrupting the tubulin, microfilaments and/or intermediate filaments of the cell. The drug may be, for example, taxon, vincristine, vinblastine, cytostatin, nocodazole, japlakinolide, latrunculin A, phalloidin, swinholide A, indanocine or myoservin.

In one aspect, the ligand is a lipid or a lipid-based molecule. The lipid or lipid-based molecule is preferably associated with a serum protein such as human serum albumin (HSA). HAS binding ligands allow distribution of the conjugates to target tissues such as liver tissue, including parenchymal cells of the liver. Other molecules capable of binding to HAS can also be used as ligands. For example, neproxin or aspirin can be used. The lipid or lipid-based molecule can: (a) increase the resistance of the conjugate to degradation; (b) increase the targeting or transport to the target cell or cell membrane; and/or (c ) can be used to modulate binding to serum proteins such as HAS.

Lipid-based ligands can be used to modulate, eg, control, the binding of the conjugate to the target tissue. For example, lipids or lipid-based ligands that bind more strongly to HSA are less likely to be targeted to the kidney and thus less likely to be cleared from the body. Lipids or lipid-based ligands that bind less well to HSA can be used to target conjugates to the kidney.

In a preferred embodiment, the lipid-based ligand binds to HAS. Preferably, it binds HAS with sufficient affinity that the conjugate will distribute preferentially to non-kidney tissues. However, preferably the affinity is not so strong that HAS-ligand binding cannot be reversed.

In another aspect, the ligand is a component, such as a vitamin or nutrient, that can be taken up by target cells, such as proliferating cells. They are very useful for the treatment of diseases characterized by, for example, undesired cell proliferation of malignant or non-malignant types such as cancer cells. Exemplary vitamins include vitamins A, E and K. Other exemplary vitamins include B vitamins such as folic acid, B12, riboflavin, biotin, pyridoxal, or other vitamins or nutrients absorbed by cancer cells.

In another aspect, the ligand is a cell penetrant, preferably a helical cell penetrant. Preferably the agent is amphoteric. Exemplary agents are peptides such as Tat peptide or antennapedia. If the agent is a peptide, it can be modified, including peptidyl mimetics, invertomers, non-peptide or pseudo-peptide linkers, and the use of D-amino acids. The helical agent is preferably an alpha helical agent, which preferably has a lipophilic phase and a lipophobic phase.

5'-Phosphate modification

In preferred embodiments, the iRNA agent is 5' phosphorylated or includes a phosphoryl analog at the 5' start. The 5'-phosphate modification of the antisense strand includes modifications consistent with RISC-mediated gene silencing. Suitable modifications include: 5'-monophosphate ((HO)2(O)P-O-5'); 5'-bisphosphate ((HO)2(O)P-O-P(HO)(O)-O-5 '); 5'-triphosphate ((HO)2(O)P-O-(HO)(O)P-O-P(HO)(O)-O-5'); 5'-guanosine cap (7-methyl or unmethylated) (7m-G-O-5'-(HO)(O)P-O-(HO)(O)P-O-P(HO)(O)-O-5'); 5'-adenosine cap ( Appp), and any modified or unmodified nucleotide cap structure (N-O-5'-(HO)(O)P-O-(HO)(O)P-O-P(HO)(O)-O-5'); 5'-monophosphorothioate (phosphothiol; (HO)2(S)P-O-5'); 5'-monodithiophosphate (phosphodithiol; (HO)(HS)( S)P-O-5'), 5'-phosphothiol esters ((HO)2(O)P-S-5'); additional combinations of any oxygen/sulfur substituted monophosphates, diphosphates and triphosphates ( e.g.5'-α-thiotriphosphate, 5'-γ-thiotriphosphate, etc.), 5'-phosphoramidate ((HO)2(O)P-NH-5', (HO)( NH2)(O)P-O-5'), 5'-phosphonic acid alkyl ester (R = alkyl = methyl, ethyl, isopropyl, propyl, etc., such as RP(OH)(O)-O- 5'-, (OH)2(O)P-5'-CH2-), 5'-alkyl ether phosphate (R = alkyl ether = methoxymethyl (MeOCH2-), ethoxymethyl , etc., such as RP(OH)(O)-O-5'-).

The sense strand can be modified to inactivate the sense strand to inhibit active RISC formation, thereby potentially reducing off-target effects. This can be accomplished by preventing 5'-phosphorylation of the sense strand, such as by modification with 5'-O-methyl ribonucleotides (see Nykanen et al., (2001) ATP requirements and small interfering RNA structure in the RNA interference pathway. Cell 107, 309-321.). Other modifications that prevent phosphorylation can also be used, eg, replacing 5'-OH with H only instead of O-Me. Alternatively, a bulky group can be added to the 5'-phosphate to convert it to a phosphodiester linker.

unnatural base

Nitropyrrolyl and nitroindolyl are a class of unnatural bases in known universal bases. A universal base is a compound that can replace any of the four naturally occurring bases without substantially affecting the melting behavior and activity of the oligonucleotide duplex. In contrast to stabilization, where hydrogen bonding is associated with naturally occurring bases, it is postulated that oligonucleotide duplexes containing 3-nitropyrrolyl bases are stabilized only by stacking. The lack of significant hydrogen bonding with the nitropyrrolyl base eliminates the specificity of a particular complementary base. In addition, various reports demonstrate that 4-, 5- and 6-nitroindolyl groups have little specificity for the four natural bases. Interestingly, 5-nitroindolyl-containing oligonucleotide duplexes were more stable than the corresponding 4-nitroindolyl- and 6-nitroindolyl-containing oligonucleotides. The procedure for the preparation of 1-(2'-O-methyl-β-D-ribofuranosyl)-5-nitroindole is described in Gaubert, G.; Wengel, J. Tetrahedron Letters 2004, 45, 5629. Other universal bases that can be used in the present invention include inosine, isoxanthine, 2-aza-inosine, 7-deaza-inosine, nitroimidazolyl, nitropyrazolyl , nitrobenzimidazolyl, nitroindazolyl, aminoindolyl, pyrropyrimidinyl, and structural derivatives thereof. For a more specific discussion of nitropyrrolyl, nitroindolyl, and other universal bases mentioned above, including synthetic steps, see Vallone et al., Nucleic Acids Research, 27(17):3589-3596 (1999); Loakes etc., J.Mol.Bio., 270:426-436(1997); Loakes et al., Nucleic Acids Research, 22(20):4039-4043(1994); Oliver et al., Organic Letters, Vol.3(13):1977 -1980(2001); Amosova et al., Nucleic Acids Research, 25(10):1930-1934(1997); Loakes et al., Nucleic Acids Research, 29(12):2437-2447(2001); Bergstrom et al., J.Am .Chem.Soc., 117:1201-1209 (1995); Franchetti et al., Biorg.Med.Chem.Lett.11:67-69 (2001); and Nair et al., Nucelosides, Nucleotides & Nucleic Acids, 20(4-7 ): 735-738 (2001).

Difluorotolyl is an unnatural base as a universal base. Difluorotolyl is the isostere of the natural base thymine. Unlike thymine, difluorotolyl does not show selectivity for any of the natural bases. Other aromatic compounds which can be used in the present invention as universal bases are 4-fluoro-6-methylbenzimidazole and 4-methylbenzimidazole. In addition, the related hydrophobic isoquinolone derivatives 3-methylisoquinolonyl, 5-methylisoquinolonyl, and 3-methyl-7-propynylisoquinolone are leading to oligonucleotide duplexes with only Oligonucleotide sequences containing natural bases are slightly less stable than universal bases. Other unnatural bases contemplated by the present invention include 7-azaindolyl, 6-methyl-7-azaindolyl, imidazopyridyl, 9-methyl-imidazopyridyl, pyrrolopyrazinyl, iso Quinolone, 7-propynylisoquinolone, propynyl-7-azaindolyl, 2,4,5-trimethylphenyl, 4-methylindolyl, 4,6-dimethylindolyl Indolyl, phenyl, naphthyl, anthracenyl, phenanthryl, pyrenyl, distyryl, naphthacene, pentacene, and their structural derivatives. For a more specific discussion of difluorotolyl, 4-fluoro-6-methylbenzimidazole, 4-methylbenzimidazole, and the other unnatural bases mentioned above, including synthetic procedures, see Schweitzer et al., J.Org .Chem., 59:7238-7242(1994); Berger et al., Nucleic Acids Research, 28(15):2911-2914(2000); Moran et al., J.Am.Chem.Soc., 119:2056-2057( 1997); Morales et al., J.Am.Chem.Soc., 121:2323-2324 (1999); Guckian et al., J.Am.Chem.Soc., 118:8182-8183 (1996); Morales et al., J. Am.Chem.Soc., 122(6):1001-1007 (2000); McMinn et al., J.Am.Chem.Soc., 121:11585-11586 (1999); Guckian et al., J.Org.Chem., 63: 9652-9656 (1998); Moran et al., Proc. Natl. Acad. Sci., 94: 10506-10511 (1997); Das et al., J. Chem. ); Shibata et al., J.Chem.Soc., Perkin Trans., 1:1605-1611 (2001); Wu et al., J.Am.Chem.Soc., 122(32):7621-7632 (2000); O 'Neill et al., J.Org.Chem., 67:5869-5875 (2002); Chaudhuri et al., J.Am.Chem.Soc., 117:10434-10442 (1995); and U.S. Patent 6,218,108.

Further details of the synthesis and use of universal bases are given in co-pending PCT/US2005/025967 by the same applicant, filed July 21, 2005, the entire contents of which are hereby incorporated by reference.

Universal bases are particularly useful when trying to target a gene in an organism that exhibits variability between different strains of that organism. This is often the case in regions of the viral genome that are considered to be highly conserved. The introduction of universal bases allows the design of iRNA agents that target a large number of different virus strains, even if the virus strains differ in one or a few (eg, up to 3) nucleotide positions.

Considering the specificity and activity of iRNA reagents, universal bases are preferably included in the region where iRNA reagents are least sensitive to nucleotide mismatches. Positions 2-9 of the antisense strand of an iRNA agent have been shown to be the most sensitive to mismatches between the antisense strand and the target mRNA, and this region is referred to as the "seed-region" of the iRNA agent. Therefore, when one or several universal bases are introduced into an iRNA agent, they are preferably introduced outside the seed region.

Table IF and Table IG show iRNA reagents that contain universal bases at complementary positions on the sense and antisense strands. However, although this is a preferred embodiment of the iRNA reagent of the present invention, the effect of the universal base in the iRNA reagent is more pronounced when the universal base is present in the antisense strand. It is thus envisioned that the base at the position in the sense strand that pairs with the universal base in the antisense strand may be the universal base or any other suitable base, such as a, u, c or g. Preferably, one will test which base position in the sense strand confers the highest activity and/or selectivity to the iRNA agent. Alternatively, the selection of bases at such specific positions in the majority of target gene variants provides that the target gene variants are inhibited in expression by the iRNA in question.

Delivery of iRNA reagents into cells

Without wishing to be bound by any theory, the chemical similarity between iRNA agents that bind cholesterol and certain lipoprotein components (e.g., cholesterol, cholesteryl esters, phospholipids) may lead to an association between iRNA agents and lipoproteins (e.g., LDL, HDL) in blood. Binding, and/or causing the iRNA agent to interact with cellular components that have an affinity for cholesterol, such as components in the cholesterol transport pathway. Lipoproteins and their constituents are taken up or processed by cells through various active or passive transport mechanisms, such as unrestricted, endocytosis of LDL-receptor-bound LDL, oxidative activity through interaction with Scavenger receptor A or other modified LDL endocytosis, Scavenger receptor B1-mediated intrahepatic HDL cholesterol uptake, pinocytosis or use of ABC (ATP binding cassette) transporters such as ABC-A1, ABC-G1 or ABC-G4 transport of cholesterol across membranes. Therefore, an iRNA agent that binds cholesterol facilitates its uptake by cells with such a transport mechanism, such as hepatocytes. Thus, the present invention provides a proof-of-concept and general method for targeting iRNA agents to cells expressing certain cell surface components, such as receptors, by binding natural ligands for said components (such as cholesterol) to iRNA agents, or By binding a chemical substance (eg cholesterol) to an iRNA agent bound to the natural ligand of the component (eg LDL, HDL).

other implementations

RNA, such as iRNA agents, can be produced within a cell, for example, by delivering an exogenous DNA template into the cell. For example, a DNA template can be inserted into a vector and used as a gene therapy vector. Gene therapy vectors can be delivered to a subject by intravenous injection, topical administration (U.S. Patent No. 5,328,470), or stereotaxic injection (see Chen et al., Proc. Natl. Acad. Sci. USA 91:3054-3057, 1994). Pharmaceutical formulations of gene therapy vectors include the gene therapy vector in an acceptable diluent, or contain a sustained release matrix in which the gene delivery vector is embedded. The DNA template, for example, comprises two transcription units, one producing a transcript comprising the upstream strand of the iRNA agent and the other producing a transcript comprising the downstream strand of the iRNA agent. When the template is transcribed, iRNA agents are produced and processed into siRNA agent fragments that mediate gene silencing.

preparation

The invention also includes pharmaceutical compositions and formulations comprising the dsRNA compounds of the invention. The pharmaceutical compositions of the present invention may be administered in a variety of ways, depending on whether local or systemic treatment is desired and depending on the area to be treated. Administration can be topical, pulmonary such as by inhalation or spraying powders or aerosols, including by spray; respiratory, intranasal, epidermal and dermal, oral or parenteral. Parenteral administration includes intravenous, intraarterial, subcutaneous, intraperitoneal or intramuscular injection or infusion; or intracranial administration, eg, intrathecal or intraventricular.

Pharmaceutical compositions and formulations for external use may include transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays, liquids or powders. Conventional pharmaceutical carriers, water, powder or oil bases, thickeners and the like are required. Coated condoms, gloves, etc. are also useful. Preferred topical formulations include those in which the dsRNAs of the present invention are mixed with topical delivery agents such as lipids, liposomes, fatty acids, fatty acid esters, steroids, chelating agents, and surfactants. Preferred lipids and liposomes include neutral (e.g. dioleoylphosphatidylethanolamine=DOPE, dimyristoylphosphatidylcholine=DMPC, distearoylphosphatidylcholine) negatively charged (e.g. dimyristoylphosphatidylcholine) acylphosphatidylglycerol = DMPG) and cationic (eg dioleoyltetramethylaminopropyl = DOTAP and dioleoylphosphatidylethanolamine = DOTMA). The DsRNAs of the invention may be encapsulated in or complexed with liposomes, particularly cationic liposomes. Alternatively, dsRNAs can be complexed with lipids, particularly cationic lipids. Preferred fatty acids and esters include, but are not limited to, arachidonic acid, oleic acid, arachidic acid, lauric acid, caprylic acid, capric acid, myristic acid, palmitic acid, stearic acid, linoleic acid, linolenic acid, dicaprate , tricaprate, glyceryl monooleate, glyceryl dilaurate, glyceryl 1-monocaprate, 1-dodecylazepan-2-one, acylcarnitine, acylcholine, or C _1-10 alkyl esters (such as isopropyl myristate IPM), monoglycerides, diglycerides or pharmaceutically acceptable salts thereof. Topical formulations are described in US Patent Application Serial No. 09/315,298, filed May 20, 1999, the entire contents of which are incorporated herein by reference.

Oral compositions and formulations include powders or granules, microparticles, nanoparticles, suspensions or solutions in aqueous or non-aqueous media, capsules, gel capsules, sachets, tablets or minitablets. There may be thickeners, flavoring agents, diluents, emulsifiers, dispersing aids or binders. Preferred oral formulations are those in which the dsRNAs of the invention are administered in combination with one or more penetration enhancers, surfactants and chelating agents. Preferred surfactants include fatty acids and/or esters or salts thereof, bile acids and/or salts thereof. Preferred bile acids/salts include chenodeoxycholic acid (CDCA) and ursodeoxycholic acid (UDCA), cholic acid, dehydrocholic acid, deoxycholic acid, glutaminocholic acid, glycocholic acid, glycosodeoxycholic acid, Taurocholic acid, taurodeoxycholic acid, taurine-24,25-dihydro-fusidate sodium and glycodihydrofusidate sodium. Preferred fatty acids include arachidonic acid, undecanoic acid, oleic acid, lauric acid, caprylic acid, capric acid, myristic acid, palmitic acid, stearic acid, linoleic acid, linolenic acid, dicaprate, tricaprate Glyceryl Monooleate, Glyceryl Dilaurate, Glyceryl 1-Monocaprate, 1-Dodecylazepan-2-one, Acylcarnitine, Acylcholine, or Monoglycerides , a diglyceride or a pharmaceutically acceptable salt (eg sodium salt) thereof. Preferred are combinations of penetration enhancers, such as combinations of fatty acids/salts and bile acids/salts. A particularly preferred combination is the sodium salt of lauric acid, capric acid and UDCA. Other penetration enhancers include polyoxyethylene-9-lauryl ester, polyoxyethylene-20-cetyl ester. The dsRNAs of the invention can be delivered orally in particulate form comprising spray-dried particles or complexed to form micro- or nanoparticles. DsRNA complexing agents include poly-amino acids; polyimines; polyacrylates; polyalkylacrylates, polyethylene oxides, polyalkylcyanoacrylates; cationized gelatin, albumin, starch, acrylates, poly Ethylene glycol (PEG) and starch; polyalkylcyanoacrylates; DEAE-derived polyimines, pollulans, cellulose and starch. Particularly preferred complexing agents include chitosan, N-trimethylchitosan, poly-L-lysine, polyhistidine, polyornithine, polyspermine, protamine, polyvinylpyrimidine , polythiodiethylaminomethylethylene P (TDAE), polyaminostyrene (e.g. p-amino), poly(methyl cyanoacrylate), poly(ethyl cyanoacrylate), poly(cyanoacrylic acid butyl), poly(isobutyl cyanoacrylate), poly(isohexyl cyanoacrylate), DEAE-methacrylate, DEAE-hexyl acrylate, DEAE-acrylamide, DEAE-albumin and DEAE-dextran Sugars, polymethylacrylate, polyhexylacrylate, poly(D,L-lactic acid), poly(DL-lactic-co-glycolic acid (PLGA), alginate, and polyethylene glycol (PEG). dsRNAs Oral formulations and preparations thereof are described in U.S. Patent Applications Ser. No. 08/886,829 (filed July 1, 1997), Ser. No. 09/108,673 (filed July 1, 1998), Ser. No. 09/256,515 (filed February 23, 1999), Ser. No. 09/082,624 (filed May 21, 1998) and Ser. No. 09/315,298 (filed May 20, 1999), hereby adopted This reference is incorporated herein by reference in its entirety.

Compositions and formulations for parenteral, intrathecal or intraventricular administration may include sterile aqueous solutions containing buffers, diluents or other suitable additives such as, but not limited to, penetration enhancers, carrier compounds and Other pharmaceutically acceptable carriers or excipients.

Pharmaceutical compositions of the present invention include, but are not limited to, solutions, emulsions, and liposome-containing formulations. These compositions can be formed from a variety of components including, but not limited to, preformed liquids, self-emulsifying solids, and self-emulsifying semi-solids.

The pharmaceutical formulations of the invention, conventionally presented in unit dosage form, may be prepared according to conventional techniques known in the pharmaceutical industry. These techniques include the step of bringing into association the active ingredient with a pharmaceutical carrier or excipient. In general, the formulations are prepared by uniformly and intimately bringing into association the active ingredient with liquid carriers or finely divided solid carriers or both, and then, if necessary, shaping the product.

The compositions of the present invention may be formulated into any possible dosage form such as, but not limited to, tablets, capsules, gel capsules, syrups, soft gels, suppositories and enemas. The compositions of the invention may also be formulated as suspensions in aqueous, non-aqueous or mixed media. Aqueous suspensions may also contain substances which increase the viscosity of the suspension including, for example, sodium carboxymethyl cellulose, sorbitol and/or dextran. The suspension may also contain stabilizers.

In one embodiment of the invention, the pharmaceutical composition may be formulated or used in the form of a foam. Pharmaceutical foams include formulations such as, but not limited to, emulsions, microemulsions, creams, gels and liposomes. Although substantially similar in nature, these formulations vary in the composition and consistency of the final product. The preparation of such compositions and formulations is well known to those skilled in the fields of pharmacy and formulation and may be used in the formulation of the compositions of the present invention.

lotion

The compositions of the invention may be prepared or formulated as emulsions. Emulsions are heterogeneous systems in which one liquid is dispersed in another as droplets larger than 0.1 μm in diameter (Idson, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., Vol. 1, p. 199; Rosoff, in Pharmaceutical Dosage Forms, Lieberman, Rieger, and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., Vol. 1, p. 245; Block in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., Vol. 2, p. 335; Higuchi et al., in Remington's Pharmaceutical Sciences, Mack Publishing Co. ., Easton, Pa., 1985, p. 301). Emulsions are often binary systems comprising two immiscible liquid phases that are intimately mixed and dispersed in one another. In general, emulsions can be of the water-in-oil (w/o) or oil-in-water (o/w) type. When the water phase is finely divided and dispersed as small droplets in the bulk oil phase, the resulting composition is called a water-in-oil (w/o) emulsion. Alternatively, when the oil is finely divided and dispersed as droplets in a large aqueous phase, the resulting composition is called an oil-in-water (o/w) emulsion. Emulsions may include other components and active drug in addition to a dispersed phase, which may be a solution in an aqueous phase, an oily phase, or itself a separate phase. Pharmaceutical excipients such as emulsifiers, stabilizers, dyes and antioxidants may also be present in the emulsions, as desired. Pharmaceutical emulsions may also be multiple emulsions comprising more than two phases, such as in the case of oil-water-oil (o/w/o) and water-oil-water (w/o/w) emulsions. Such complex formulations often have advantages over simple binary emulsions. In a multiple emulsion, individual oil droplets of an o/w emulsion surround small water droplets to form a w/o/w emulsion. Also a system containing oil droplets within water droplets stabilized in an oil continuous phase is an o/w/o emulsion.

Emulsions are characterized by thermodynamic instability. Typically, the dispersed or discontinuous phase of an emulsion is well dispersed in the external or continuous phase and is maintained in this form by the emulsifier or the viscosity of the formulation. The phase of an emulsion can be semi-solid or solid, like an emulsion salve or cream. Other methods of stabilizing emulsions require the use of emulsifiers, which can be added to either phase of the emulsion. Emulsifiers can be broadly divided into 4 classes: synthetic surfactants, natural emulsifiers, absorbent matrices and finely divided solids (Idson, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., Vol. 1, p. 199).

Synthetic surfactants, also known as surfactants, are widely used in emulsion formulations and are reviewed in the literature (Rieger, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc. ., New York, N.Y., volume 1, p.285; Idson, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), Marcel Dekker, Inc., New York, N.Y., 1988, volume 1, p.199) . Surfactants are generally amphoteric and include hydrophilic and hydrophobic moieties. The ratio of the hydrophilic and hydrophobic properties of a surfactant is defined as the hydrophilic/lipophilic balance (HLB), which is an effective tool for the classification and selection of surfactants in the manufacture of formulations. Surfactants can be divided into different types according to the nature of the hydrophilic group: nonionic, anionic, cationic and amphoteric (Rieger, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker , Inc., New York, N.Y., Vol. 1, p. 285).

Natural emulsifiers used in emulsion formulations include lanolin, beeswax, phospholipids, lecithin and acacia. Absorbent bases are hydrophilic in that they absorb water to form a w/o emulsion but maintain a semi-solid consistency, such as anhydrous lanolin and hydrophilic petrolatum. Finely divided solids can also be used as good emulsifiers, especially in combination with surfactants and in viscous formulations. These solids include polar inorganic solids such as heavy metal hydroxides, non-intumescent ceramics such as bentonite, attapulgite, hectorite, kaolin, montmorillonite, colloidal aluminum silicate and colloidal magnesium aluminum silicate, pigments and non-polar solids such as carbon or tristearin.

Emulsion formulations also include a wide variety of non-emulsifying materials whose presence contributes to the properties of the emulsion. These non-emulsifying materials include fats, oils, waxes, fatty acids, fatty alcohols, fatty acid esters, humectants, hydrocolloids, preservatives, and antioxidants (Block, in Pharmaceutical Dosage Forms, Lieberman, Rieger, and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., Vol. 1, p. 335; Idson, in Pharmaceutical Dosage Forms, Lieberman, Rieger, and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y. , Vol. 1, p. 199).

Hydrocolloids or hydrocolloids include natural rubber and synthetic polymers such as polysaccharides (e.g., gum arabic, agar, alginic acid, carrageenan, guar gum, karaya, and tragacanth), cellulose-derived substances (for example, carboxymethylcellulose and carboxypropylcellulose), and synthetic polymers (for example, carbomers, cellulose ethers, and carboxyvinyl polymers). These disperse or swell in water to form a colloidal solution, which stabilizes the emulsion by forming a strong interfacial film around the dispersed phase liquid and by increasing the viscosity of the external phase.

Since emulsions often contain significant amounts of components that support microbial growth, such as carbohydrates, proteins, sterols, and phospholipids, these formulations often contain preservatives. Commonly used preservatives in emulsion formulations include methylparaben, propylparaben, quaternary ammonium salts, benzalkonium chloride, p-hydroxybenzoate, and boric acid. Antioxidants are often added to emulsion formulations to prevent deterioration of the formulation. Antioxidants used may be free radical scavengers such as tocopherol, alkyl gallate, butylated hydroxyanisole, butylated hydroxytoluene or reducing agents such as ascorbic acid and sodium metabisulfite, and antioxidant synergists such as citric acid, tartaric acid, and lecithin.

The use of emulsion formulations by dermal, oral and parenteral routes and their methods of manufacture have been reviewed in the literature (Idson, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., Vol. 1, p. 199). Emulsion formulations for oral delivery have been widely used due to the convenience of formulation and the high efficiency of absorption and bioavailability (Rosoff, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p.245; Idson, in Pharmaceutical Dosage Forms, Lieberman, Rieger, and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p.199). Mineral oil-based laxatives, oil-soluble vitamins, and high-fat nutritional preparations are also among the commonly used substances taken orally in o/w emulsions.

In one embodiment of the invention, the composition of dsRNAs and nucleic acids is formulated as a microemulsion. Microemulsions are defined as systems of water, oil, and hydrophilic lipid molecules that are individual visually homogeneous and thermodynamically stable liquid solutions (Rosoff, in Pharmaceutical Dosage Forms, Lieberman, Rieger, and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., Vol. 1, p. 245). Typically, microemulsions are prepared by first dispersing the oil in an aqueous surfactant solution and then adding a sufficient amount of the fourth component (usually a medium chain alcohol) to form a transparent system. Microemulsions are thus thermodynamically stable isotropic clear dispersions of two immiscible liquids, stabilized by an interfacial film of surface-active molecules (Leung and Shah, in: Controlled Release of Drugs: Polymers and Aggregate Systems , Rosoff, M., Ed., 1989, VCH Publishers, New York, pp. 185-215). Microemulsions are usually prepared by combining 3 to 5 components including oil, water, surfactants, co-surfactants and electrolytes. Whether a microemulsion is water-in-oil (w/o) or oil-in-water (o/w) depends on the nature of the oil and the surfactant used, and also on the structure and geometry of the polar heads and hydrocarbon tails of the surfactant molecules Geometric packing (Schott, in Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa., 1985, p. 271).

Phenomenological methods using phase diagrams have been extensively studied and those skilled in the art have acquired a comprehensive knowledge of how to formulate microemulsions (Rosoff, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p.245; Block, in Pharmaceutical Dosage Forms, Lieberman, Rieger, and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p. 335). The advantage of microemulsions over conventional emulsions is the dissolution of water-insoluble drugs in naturally occurring thermodynamically stable droplet formulations.

Surfactants used in the preparation of microemulsions include, but are not limited to, ionic surfactants, nonionic surfactants, Brij96, polyoxyethylene oleyl ether, polyglycerides of fatty acids, tetraglyceride monolaurate (ML310), mono-oil Tetraglyceryl Monooleate (MO310), Hexaglyceryl Monooleate (PO310), Hexaglyceryl Pentaoleate (PO500), Decaclyceryl Monocaprate (MCA750), Decaclyceryl Monooleate (MO750), Semi Oil Decacylglyceryl Decacylate (SO750), Decacylglyceryl Decoleate (DAO750), alone or in combination with a co-surfactant. Co-surfactants are typically short-chain alcohols such as ethanol, 1-propanol, and 1-butanol, which are used to create abnormal films by penetrating into the surfactant film and creating voids between the surfactant molecules, thereby increasing interface fluidity. However, microemulsions can be prepared without co-surfactants, and alcohol-free self-emulsifying microemulsion systems are known in the art. The aqueous phase is generally but not limited to water, aqueous drug solution, glycerin, PEG300, PEG400, polyglycerol, propylene glycol, and ethylene glycol derivatives. Oil phases include, but are not limited to, e.g. Captex 300, Captex 355, Capmul MCM, fatty acid esters, medium chain ( _C8 - _C12 ) mono-, diglycerides and triglycerides, polyoxyethyleneated fatty acid esters , fatty alcohols, PEGylated glycerides, saturated PEGylated glycerides, saturated PEGylated C ₈ -C ₁₀ glycerides, vegetable oils and silicone oils.

Microemulsions have received special attention from the perspective of drug solubilization and enhanced drug absorption. Lipid-based microemulsions (o/w and w/o) have been proposed to enhance the bioavailability of drugs, including peptides (Constantinides et al., Pharamaceutical Research, 1994, 11, 1385-1390; Ritschel, Meth. Find. Exp. Clin . Pharmacol., 1993, 13, 205). Microemulsions have the potential to improve drug solubilization, prevent drug enzymatic hydrolysis, enhance absorption possibly due to surfactant-induced changes in membrane fluidity and permeability, facilitate preparation, facilitate oral administration compared to solid dosage forms, improve clinical efficacy, and reduce Advantages of toxicity (Constantinides et al., Pharmaceutical Research, 1994, 11, 1385; Ho et al., J. Pharm. Sci., 1996, 85, 138-143). Microemulsions often form spontaneously when their components are mixed at room temperature. This is particularly advantageous when formulating thermolabile drugs, peptides or dsRNAs. Microemulsions are also useful for transdermal delivery of active ingredients in cosmetic or pharmaceutical applications. The microemulsion compositions and formulations of the invention are expected to facilitate increased systemic absorption of dsRNAs and nucleic acids from the gastrointestinal tract and improved local cellular uptake of dsRNAs and nucleic acids in the gastrointestinal tract, vagina, oral cavity, and other areas of administration.

The microemulsions of the present invention may contain additional components and additives (such as sorbitan monostearate (Grill3), caprylic/capric macrogolglycerides (Labrasol), and penetration enhancers) to improve formulation properties and enhance the uptake of the dsRNAs and nucleic acids of the invention. Penetration enhancers in the microemulsions of the present invention can be classified into one of five major categories: surfactants, fatty acids, bile salts, chelating agents, and non-chelating non-surfactants (Lee et al., Critical Reviews in Therapeutic Drug Carriers Systems , 1991, p.92). Each of the categories already discussed above.

Liposomes

In addition to microemulsions that have been studied and used in pharmaceutical formulations, there are many regular surfactant structures including monolayers, micelles, bilayers and vesicles. Vesicles such as liposomes have attracted much attention from the perspective of drug delivery due to their specificity and long-lasting effect. The term "liposome" as used herein refers to a vesicle composed of amphiphilic lipids arranged in a spherical bilayer or bilayer.

Liposomes are unilamellar or multilamellar vesicles having a membrane formed of a lipophilic material and an aqueous inner core. The aqueous portion contains the composition to be delivered. Cationic liposomes have the advantage of being able to fuse with the cell wall. Non-cationic liposomes can be taken up in vivo by macrophages, although they cannot fuse efficiently with cell walls.

To penetrate intact mammalian skin, lipid vesicles must pass through a series of pores, each less than 50 nm in diameter, subject to a suitable transdermal gradient. Therefore, there is a need to use liposomes that can be highly deformed and pass through such pores.

Other advantages of liposomes include: liposomes derived from natural phospholipids are biocompatible and biodegradable; liposomes can incorporate large amounts of water and lipid-soluble drugs; liposomes prevent encapsulation of Drugs are metabolized and degraded (Rosoff, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p.245). Important considerations in the preparation of liposome formulations are lipid surface charge, vesicle size, and aqueous volume of liposomes.

Liposomes are very useful for delivery and delivery of active substances to the site of action. Because the liposome membrane is similar to the biological membrane structure, when the liposome is used in the tissue, the liposome begins to fuse with the cell membrane, and as the liposome fuses, the cell grows, and the liposome content enters the cell. There the active ingredients take effect.

Liposome formulations have been the focus of extensive research as a delivery model for a variety of drugs. There is increasing evidence that liposomes have several advantages over other formulations for topical use. Such advantages include reduced side effects associated with high systemic absorption of administration, increased accumulation of administration on target, and the possibility of delivering multiple drugs (hydrophilic or hydrophobic) to the skin.

Several reports have detailed the effect of liposomes on delivery agents, including high molecular weight DNA, into the skin. Compounds (including analgesics, antibodies, hormones, and macromolecular DNAs) can be delivered into the skin. Most applications target the upper epidermis.

Liposomes fall into two broad categories. Cationic liposomes are positively charged liposomes that interact with negatively charged DNA molecules to form stable complexes. The positively charged DNA/liposome complex binds to the negatively charged cell surface and is internalized in endosomes. Due to the acidic pH in endosomes, liposomes rupture, releasing their contents into the cytoplasm (Wang et al., Biochem. Biophys. Res. Commun., 1987, 147, 980-985).

Liposomes are pH-sensitive or negatively charged, and capture DNA rather than complexes. Since DNA and lipids are similarly charged, repulsion rather than recombination occurs. However, some DNA is trapped within the aqueous inner core of the liposome. pH-sensitive liposomes have been used to deliver the DNA encoding the thymidine kinase gene into cell monolayers in culture. The expression of foreign genes is detected by target cells (Zhou et al., Journal of Controlled Release, 1992, 19, 269-274).

One major type of liposomal composition includes phospholipids other than naturally derived phosphatidylcholine. Neutral liposome compositions can be formed, for example, from dimyristoylphosphatidylcholine (DMPC) or dipalmitoylphosphatidylcholine (DPPC). Cationic liposome compositions are generally formed from dimyristoylphosphatidylglycerol, while cationic membranous liposomes are primarily formed from dioleoylphosphatidylethanolamine (DOPE). Another liposome composition is formed from phosphatidylcholine (PC), eg, soybean PC and egg PC. The other is formed from a mixture of phospholipids and/or phosphatidylcholines and/or cholesterol.

Several studies have evaluated the topical delivery of liposomal drug formulations to the skin. Application of liposomes containing interferon to the skin of guinea pigs resulted in a reduction in cutaneous herpes, whereas delivery of interferon by other means (e.g. as a solution or emulsion) was ineffective (Weiner et al., Journal of Drug Targeting, 1992, 2, 405- 410). Then, also another study tested the effectiveness of interferon as part of liposomal formulations relative to the use of aqueous systems to deliver interferon, and concluded that liposomal formulations are superior to aqueous systems (duPlessis et al., Antiviral Research, 1992, 18, 259- 265).

Nonionic liposome systems have proven useful in delivering drugs to the skin, especially systems containing nonionic surfactants and cholesterol. Nonionic liposome formulations contain Novasome.TM.I (glyceryl dilaurate/cholesterol/polyoxyethylene-10-stearyl) and Novasome.TM.II (glyceryl distearate/cholesterol/polyoxy Ethylene-10-stearyl ester), which was used to deliver cyclosporin-A to the dermis of mice. The results showed that such a nonionic liposome system effectively drives the deposition of cyclosporin-A to the different layers of the skin (Hu et al. S.T.P. Pharma. Sci., 1994, 4, 6, 466).

Liposomes also include "sterically stabilized" liposomes, which term refers herein to liposomes that include one or more specific lipids that, when incorporated into a liposome, differ from those lacking such specific lipids. Enhanced cycle life compared to liposomes. An example of a sterically stable liposome is a portion in the vesicle-forming lipid portion of a liposome (A) comprising one or more glycolipids, such as ganglioside G _ml , or (B) Derivatized with one or more hydrophilic polymers, such as polyethylene glycol (PEG) groups. Without being bound by any particular theory, it is believed in the art that, at least for sterically stabilized liposomes containing gangliosides, sphingomyelin, or PEG-derived lipids, these sterically stabilized liposomes enhance The circulatory half-life of ® is derived from reduced intracellular uptake of the reticuloendothelial system (RES) (Allen et al., FEBS Letters, 1987, 223, 42; Wu et al., Cancer Research, 1993, 53, 3765).

Liposomes comprising one or more of various glycolipids are known in the art. Papahadjopoulos et al. (Ann. NY Acad, Sci., 1987, 507, 64) reported the ability of ganglioside G _ml , galactoside sulfate and phosphatidylinositol to improve the blood half-life of liposomes. These findings are explained in detail by Gabizon et al. (Proc. Natl. Acad. Sci. USA, 1988, 85, 6949). US Patent No. 4,837,028 and WO 88/04924, both to Allen et al., disclose liposomes comprising (1) sphingomyelin and (2) ganglioside _Gml or galactocerebroside sulfate. US Patent No. 5,543,152 (Webb et al.) discloses liposomes comprising sphingomyelin. Liposomes comprising 1,2-sn-dimyristoylphosphorylcholine are disclosed in WO 97/13499 (Lim et al.).

Liposomes containing lipids derived from one or more hydrophilic polymers and methods for their preparation are known in the art. Sunamoto et al. (Bull. Chem. Soc. Jpn., 1980, 53, 2778) describe liposomes containing a nonionic detergent, 2C _1215G , which contain a PEG group. Illum et al. (FEBS Lett., 1984, 167, 79) showed that a polyethylene glycol hydrophilic coating of polystyrene particles resulted in a greatly increased blood half-life. Synthetic phospholipids modified by appending carboxyl groups of polyalkylene glycols such as PEG are illustrated by Sears (US Patent Nos. 4,426,330 and 4,534,899). Klibanov et al. (FEBS Lett., 1990, 268, 235) describe experiments showing that liposomes containing phosphatidylethanolamine (PE) derivatized with PEG or PEG stearate can significantly increase blood circulation half-life. Blume et al. (Biochimica et Biophysica Acta, 1990, 1029, 91) extended the above observations to another PEG-derived phospholipid, such as DSPE-PEG, formed from the combination of distearoylphosphatidylethanolamine (DSPE) and PEG. Liposomes with covalently bound PEG groups on the outer surface are described in Fisher's European Patent EP 0 445 131 B1 and WO 90/04384. PEG-derived liposome compositions containing 1-20 mol% PE and methods for their use are described in Woodle et al. (US Patent Nos. 5,013,556 and 5,356,633) and Martin et al. Explained. Liposomes containing a number of other lipid-polymer conjugates are described in WO 91/05545 and US Pat. Nos. 5,225,212 (both to Martin et al.) and WO 94/20073 (Zalipsky et al.). Liposomes containing PEG-modified ceramide lipids are described in WO 96/10391 (Choi et al). US Patent No. 5,540,935 (Miyazaki et al.) and US Patent No. 5,556,948 (Tagawa et al.) describe PEG-containing liposomes, which can be further surface derivatized with functional groups.

A small number of liposomes containing nucleic acids are known in the art. WO 96/40062 Thierry et al. disclose methods for encapsulating high molecular weight nucleic acids in liposomes. U.S. Patent No. 5,264,221 Tagawa et al. disclose protein-conjugated liposomes and assert that the contents of the liposomes may include dsRNA. U.S. Patent No. 5,665,710 Rahman et al. describe a method of encapsulating oligonucleotides in liposomes. WO 97/04787 Love et al. disclose liposomes containing dsRNAs targeting the raf gene.

Transfersomes are another type of liposome and are highly deformable lipid aggregates that are important candidates as drug delivery vehicles. Transfersomes can be described as lipid liquids that are highly deformable allowing them to readily pass through pores smaller than liquid droplets. Transfersomes adapt to their environment of use, eg they can self-optimize (adapt to the shape of the pores in the skin), self-heal, often reach their target without cleavage, and often self-loading. To enable transfersomes, surface edge-activating agents, usually surfactants, are added to standard liposome compositions. Transfersomes have been used to deliver serum albumin to the skin. Transfersome-mediated delivery of serum albumin was shown to be as effective as subcutaneous injection of serum albumin-containing solutions.

Surfactants find wide application in formulations such as emulsions (including microemulsions) and liposomes. The most common method of classification and property ranking of the many different classes of surfactants (natural or synthetic) utilizes the Hydrophile/Lipophile Balance (HLB). The nature of the hydrophilic group (also known as the "head") is the most useful tool for classifying different surfactants used in formulations. (Rieger, in Pharmaceutical Dosage Forms, Marcel Dekker, Inc., New York, N.Y., 1988, p. 285).

If a surfactant molecule cannot be ionized, it is classified as a nonionic surfactant. Nonionic surfactants are widely used in pharmaceuticals and cosmetics and are available over a wide pH range. Typically their HLB values range from 2 to about 18, depending on their structure. Nonionic surfactants include nonionic esters such as ethylene glycol esters, propylene glycol esters, glycerol esters, polyglyceryl esters, sorbitan esters, sucrose esters, and ethoxylated esters. Nonionic alkyl amides and ethers such as fatty alcohol ethoxylates, propoxylated alcohols and ethoxylated/propoxylated chimeric polymers are also in this class. Polyoxyethylene surfactants are the most commonly used nonionic surfactants.

If the surfactant molecules are negatively charged when dissolved or dispersed in water, the surfactant is classified as an anionic surfactant. Anionic surfactants include carboxylates such as fatty acid salts, acyl lactylates, amides of amino acids, sulfates such as alkyl sulfates and ethoxylated alkyl sulfates, sulfonates such as alkyl benzenesulfonate, acyl Isethionates, acyl taurates and phosphates. The most important anionic surfactants are alkyl sulfates and fatty acid salts.

A surfactant is classified as a cationic surfactant if its molecules are positively charged when dissolved or dispersed in water. Cationic surfactants include quaternary ammonium salts and ethoxylated amines. The most commonly used of these are quaternary ammonium salts.

If the surfactant molecule can be positively or negatively charged, the surfactant is classified as an amphoteric surfactant. Amphoteric surfactants include acrylic acid derivatives, substituted alkyl amides, N-alkyl betaines and phospholipids.

The use of surfactants in pharmaceutical products, formulations and emulsions is reviewed in (Rieger, in Pharmaceutical Dosage Forms, Marcel Dekker, Inc., New York, N.Y., 1988, p.285).

penetration enhancer

In one embodiment, the present invention utilizes various penetration enhancers to achieve efficient delivery of nucleic acids, particularly dsRNAs, to the skin of animals. Most drugs exist in solution in ionized or non-ionized form. Usually, however, only lipid-soluble or lipophilic drugs cross cell membranes. It was found that even non-lipophilic drugs can cross cell membranes if the membrane to cross is treated with a penetration enhancer. In addition to facilitating the penetration of non-lipophilic drugs across cell membranes, penetration enhancers also enhance the permeability of lipophilic drugs.

Penetration enhancers can be classified into one of five broad categories, namely surfactants, fatty acids, bile salts, chelating agents, and non-chelating non-surfactants (Lee et al., Critical Reviews in Therapeutic Drug Carriers Systems, 1991, p. 92 ). Each of the above categories of penetration enhancers will be described in detail below.

Surfactant: In relation to the present invention, a surfactant (or "surfactant") is a chemical substance that, when dissolved in an aqueous solution, lowers the surface tension of the solution or lowers the interfacial tension between the aqueous solution and other liquids, which can enhance the passage of dsRNAs Absorption by mucous membranes. In addition to bile salts and fatty acids, these penetration enhancers include, for example, sodium laurate sulfate, polyoxyethylene-9-lauryl and polyoxyethylene-20-hexadecyl esters) (Lee et al., Critical Reviews in Therapeutic Drug Carriers Systems, 1991, p.92); and perfluorinated chemical emulsions, such as FC-43 (Takahashi et al., J.Pharm.Pharmacol., 1988, 40, 252).

Fatty acids: Various fatty acids and their derivatives that act as penetration enhancers include, for example, oleic acid, lauric acid, capric acid (n-capric acid), myristic acid, palmitic acid, stearic acid, linoleic acid, linolenic acid , Dicaprate, Tricaprate, Glyceryl Monooleate (1-Monoleoyl-rac-glycerol), Glyceryl Dilaurate, Caprylic Acid, Arachidonic Acid, Glyceryl 1-Monocaprate, 1 - dodecylazepan-2-ones, acylcarnitines, acylcholines, their C ₁ -C ₁₀ alkyl esters (for example, methyl, isopropyl and t-butyl esters), and their mono -glycerides and di-glycerides (e.g., oleate, laurate, caprate, myristate, palmitate, stearate, linoleate, etc.) (Lee et al., Critical Reviews in Therapeutic Drug Carrier Systems, 1991, p.92; Muranishi, Critical Reviews in Therapeutic Drug Carriers Systems, 1990, 7, 1-33; El Hariri et al., J.Pharm.Pharmacol., 1992, 44, 651-654).

Bile salts: The physiological roles of bile include facilitating the dispersion and absorption of lipids and fat-soluble vitamins (Brunton, Chapter 38 in: Goodman & Gilman's The Pharmacological Basis of Therapeutics, 9th Ed., Hardman et al. Eds., McGraw-Hill, New York , 1996, pp.934-935). Various natural bile salts and their synthetic derivatives act as penetration enhancers. Thus, the term "bile salts" includes any natural component of bile and any synthetic derivatives thereof. The bile salts of the present invention include, for example, cholic acid (or its pharmaceutically acceptable sodium salt, sodium cholate), dehydrocholic acid (sodium dehydrocholate), deoxycholic acid (sodium deoxycholate), glutamine Cholic Acid (Sodium Glutaminocholate), Glycocholic Acid (Sodium Glycocholate), Glycodeoxycholic Acid (Sodium Glycodeoxycholate), Taurocholic Acid (Sodium Taurocholate), Taurodeoxycholic Acid Acid (Sodium Taurodeoxycholate), Chenodeoxycholic Acid (Sodium Chenodeoxycholate), Ursodeoxycholic Acid (UDCA), Sodium Taurine-24,25-Dihydro-Fusidate (STDHF), Sugar Sodium fusidate and polyoxyethylene-9-lauryl ester (POE) (Lee et al., Critical Reviews in Therapeutic Drug Carriers Systems, 1991, page 92; Swinyard, Chapter 39 In: Remington's Pharmaceutical Sciences, 18th Ed., Gennaro, ed., MackPublishing Co., Easton, Pa., 1990, pages 782-783; Muranishi, Critical Reviews in Therapeutic Drug Carriers Systems, 1990, 7, 1-33; Yamamoto et al., J.Pharm.Exp.Ther., 1992, 263, 25; Yamashita et al., J. Pharm. Sci., 1990, 79, 579-583).

Chelating Agents: Chelating agents in relation to the present invention can be defined as compounds that remove metal ions from solution by forming complexes with them, which can enhance the absorption of dsRNAs through mucous membranes. Considering its use as a penetration enhancer in the present invention, a chelating agent has the added advantage of being a DNase inhibitor, since most characterized DNA nucleases require divalent metal catalysis and thus can be inhibited by chelating agents (Jarrett, J. Chromatogr., 1993, 618, 315-339). Chelating agents of the present invention include, but are not limited to, disodium ethylenediaminetetraacetic acid (EDTA), citric acid, salicylates (e.g., sodium salicylate, sodium 5-methoxysalicylate, and homovanilate). )), N-acyl derivatives of collagen, N-aminoacyl derivatives (enamines) of alkyl polyoxyethylene ether-9 and β-diketones (Lee et al., Critical Reviews in Therapeutic Drug Carriers Systems, 1991, page 92 ; Muranishi, Critical Reviews in Therapeutic Drug Carriers Systems, 1990, 7, 1-33; Buur et al., J. Control Rel., 1990, 14, 43-51).

Non-chelating non-surfactant: A non-chelating non-surfactant penetration enhancing compound as used herein can be defined as a compound that does not exhibit significant activity of chelating agents or surfactants but enhances the absorption of dsRNA through the digestive mucosa (Muranishi, Critical Reviews in Therapeutic Drug Carriers Systems, 1990, 7, 1-33). Such penetration enhancers include, for example, unsaturated cyclic urea, 1-alkyl- and 1-alkynyl azacyclo-aliphatic ketone derivatives (Lee et al., Critical Reviews in Therapeutic Drug Carriers Systems, 1991, page 92); and Non-steroidal anti-inflammatory agents such as diclofenac sodium, indomethacin and phenylbutazone (Yamashita et al., J. Pharm. Pharmacol., 1987, 39, 621-626).

Agents that enhance the uptake of dsRNAs at the cellular level may also be added to the pharmaceutical and other compositions of the invention. For example, known are cationic lipids, such as liposomes (Junichi et al, U.S. Patent No. 5,705,188), cationic glycerol derivatives, and polycationic molecules, such as polylysine (Lollo et al, PCT Application WO 97/30731) , can also enhance the cellular uptake of dsRNAs.

Other agents can be used to enhance penetration of administered nucleic acids, including glycols, such as ethylene glycol and propylene glycol, pyrroles, such as 2-pyrrole, azone, and terpenes, such as limonene and menthone.

carrier

Certain compositions of the invention also include a carrier compound in the formulation. As used herein, a "carrier compound" or "carrier" may refer to a nucleic acid or analog thereof that is inert (i.e., not biologically active per se), but is recognized as a nucleic acid during in vivo processes, for example by degrading a biologically active nucleic acid or promoting Their exclusion from circulation reduces the bioavailability of biologically active nucleic acids. Co-administration of nucleic acid and carrier compound, generally in excess of the latter, can significantly reduce the amount of nucleic acid recovered in the liver, kidney or other extracirculatory reservoirs, presumably because the carrier compound competes with the nucleic acid for general receptors. For example, in combination with polyinosinic acid, dextran sulfate, polycytidic acid, or 4-acetamido-4'isothiocyanato-stilbene-2,2'-disulfonic acid When administered, the recovery of phosphorothioate dsRNA in liver tissue can be partially reduced (Miyao et al., Antisense Res.Dev., 1995, 5, 115-121; Takakura et al., Antisense & Nucl.Acid Drug Dev., 1996, 6, 177-183.).

excipient

In contrast to carrier compounds, a "pharmaceutical carrier" or "excipient" is a pharmaceutically acceptable solvent, suspending agent, or any pharmacologically inert vehicle for delivering one or more nucleic acids to an animal. Excipients can be liquid or solid, and are selected with regard to the form of administration intended to bring about the desired volume, consistency, etc. when mixed with the nucleic acid and other components of the pharmaceutical composition administered. Typical pharmaceutical carriers include, but are not limited to, binders (e.g., pregelatinized cornstarch, polyvinylpyrrolidone or hydroxypropylmethylcellulose, etc.); fillers (e.g., lactose or other sugars, microcrystalline cellulose , pectin, gelatin, calcium sulfate, ethyl cellulose, polyacrylate or calcium hydrogen phosphate, etc.); lubricants (such as magnesium stearate, talc, quartz, colloidal silicon dioxide, stearic acid, stearic acid metal salts, hydrogenated vegetable oils, corn starch, polyethylene glycol, sodium benzoate, sodium acetate, etc.); decomposers (such as starch, sodium starch glycolate, etc.); and humectants (such as sodium lauryl sulfate, etc.).

Pharmaceutically acceptable organic or inorganic excipients suitable for non-parenteral administration that have no adverse reaction with nucleic acids can be used to formulate the compositions of the present invention. Suitable pharmaceutically acceptable carriers include, but are not limited to, water, saline solution, alcohol, polyethylene glycol, gelatin, lactose, amylose, magnesium stearate, talc, silicic acid, viscose, hydroxymethylcellulose, polyvinylpyrrolidone, etc.

Topical formulations of nucleic acids may include sterile and non-sterile aqueous solutions, non-aqueous solutions in common solvents such as alcohols, or solutions of nucleic acids in liquid or solid oily phases. Solutions may contain buffers, diluents and other suitable additives. Pharmaceutically acceptable organic or inorganic excipients suitable for non-parenteral administration that have no adverse reaction with nucleic acids can also be used.

Suitable pharmaceutically acceptable excipients include, but are not limited to, water, saline solution, alcohol, polyethylene glycol, gelatin, lactose, amylose, magnesium stearate, talc, silicic acid, viscose, hydroxymethylcellulose Polyvinylpyrrolidone, polyvinylpyrrolidone, etc.

Pharmaceutical compositions suitable for delivery to the respiratory tract

Another aspect of the invention provides for the delivery of IRNA agents to the respiratory tract, particularly for the treatment of cystic fibrosis. The respiratory tract consists of the upper airway (including the oropharynx and larynx) then the lower airway (including the trachea, which then bifurcates into the bronchi and bronchioles. The upper and lower airways are called conductive airways. The terminal bronchioli divide into respiratory bronchioli , and then reach the final respiratory zone, the alveoli or the deep lung. The lung or alveoli are the primary target for inhaled therapeutic aerosols for systemic delivery of iRNA agents.

Compositions for pulmonary delivery can be delivered by inhalation of the dispersion by the patient so that the composition, preferably the iRNA agent, in the dispersion can reach the lungs, where it can pass through the alveoli directly into the blood circulation. Pulmonary delivery is effective for both systemic delivery and local delivery for the treatment of pulmonary diseases.

Pulmonary delivery can be achieved by different methods, including the use of nebulized, aerosolized micellar and dry powder based formulations; inhalation administration can be oral and/or nasal. Delivery can be achieved with liquid sprays, aerosol-based inhalants, and dry powder dispersion devices. Metered-dosed devices are preferred. One of the benefits of using a nebulizer or inhaler is that contamination is minimized because the device is self-contained. Dry powder dispensing devices deliver drugs that are readily formulated as dry powders. The iRNA composition can be stored stably as a lyophilized or spray-dried powder by itself or in combination with a suitable powder carrier. Delivery of the inhalation composition may be mediated by a quantitative timing element including a timer, doser, time measuring device or time indicator incorporated into the device to allow tracking of dosage, monitoring of the patient during aerosol drug administration. Adaptation, and/or dose triggering.

Examples of pharmaceutical devices for aerosol delivery include dose-measured inhalers (MDIs), dry powder inhalers (DPIs), and air-jet nebulizers. Exemplary inhalation delivery systems that can be used to deliver iRNA agents of the invention are described, for example, in U.S. Patent Nos. 5,756,353; . Other aerosol formulations that can be used to deliver iRNA agents are described in US Patent Nos. 6,294,153; 6,344,194; 6,071,497, and PCT applications WO02/066078; WO02/053190; WO01/60420; WO00/66206. Furthermore, methods for delivering iRNA agents can be based on methods used in inhalational delivery of other oligonucleotides (such as antisense oligonucleotides), such as in Templin et al., Antisense Nucleic Acid Drug Dev, 2000, 10:359-68; Sandrasagra et al., Expert Opin Biol Ther, 2001, 1:979-83; Sandrasagra et al., Antisense Nucleic Acid Drug Dev, 2002, 12:177-81.

Delivery of agents of the invention may also include the administration of so-called "prodrugs," i.e., formulations or chemical modifications of therapeutic substances that require some form of processing or systemic transport native to the target organism for release, preferably at the desired Site-of-action release; the latter embodiment can be used in conjunction with respiratory tract delivery, as well as with other embodiments of the invention. For example, the human lungs can remove or rapidly degrade hydrolytically fragmented deposited aerosols within minutes to hours. In the upper airway, ciliated epithelial cells facilitate the "mucociliary excalator" by which particles are swept from the airway into the mouth. Pavia, D., "Lung Mucociliary Clearance," in Aerosols and the Lung: Clinical and Experimental Aspects, Clarke, S.W. and Pavia, D., Eds., Butterworths, London, 1984. Inside the lung, alveolar macrophages can phagocytose particles as they are deposited. Warheit et al. Microscopy Res. Tech., 26:412-422 (1993); and Brain, J.D., "Physiology and Pathophysiology of Pulmonary Macrophages," in The Reticuloendothelial, S.M. Reichard and J. Filkins, Eds., Plenum, New. York. , pp. 315-327, 1985.

In preferred embodiments, particularly where systemic administration of the iRNA agent is desired, the aerosolized iRNA agent is made into microparticles. Microparticles with a diameter of 0.5-10 μm can penetrate the lungs and pass through most natural barriers. A diameter of less than 10 μm is required to bypass the larynx; a diameter of 0.5 μm or more is required to avoid exhalation.

other components

The compositions of the present invention also contain other adjuvant components conventional in pharmaceutical compositions at levels established in the art. Thus, for example, the composition may contain additional, compatible, pharmaceutically active substances, for example, antipruritic, astringent, local anesthetic or anti-inflammatory agents, or may contain various dosage forms for physically formulating the compositions of the invention other substances such as dyes, flavorings, preservatives, antioxidants, opacifiers, thickeners and stabilizers. However, when added, such materials should not unduly affect the biological activity of the ingredients of the compositions of the present invention. The preparation can be sterilized and, if desired, mixed with auxiliary agents, such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts, so as to affect the osmotic pressure, buffer, color, taste and / or not with the preparation Nucleic acids have adverse interactions with aromatic substances, etc.

Aqueous suspensions may contain substances which increase the viscosity of the suspension including, for example, sodium carboxymethyl cellulose, sorbitol and/or dextran. The suspension may also contain stabilizers.

Certain embodiments of the invention provide a pharmaceutical composition comprising (a) one or more dsRNA agents and (b) one or more other chemotherapeutic agents that act by mechanisms other than RNA interference. Examples of such chemotherapeutic agents include, but are not limited to, daunorubicin, daunorubicin, dactinomycin, doxorubicin, epirubicin, idarubicin, 4'-deoxydoxorubicin (esorubicin), erythromycin, Lymycin, Mafosfamide, Ifosfamide, Cytarabine, Bis-Chloroethylnitrourea, Busulfan, Mitomycin C, Actinomycin D, Mithramycin, Mithramycin Hydroxyprogesterone, Testosterone, Tamoxifen, Dacarbazine, Procarbazine, Hexamethylmelamine, Pentamethylmelamine, Mitoxantrone, Amsacridine, Chlorambucil, Methylcycline Hexylnitrourea, nitrogen mustard, phenylalanine mustard, cyclophosphamide, 6-mercaptopurine, 6-thioguanine, cytarabine, 5-azacytidine, hydroxyurea, deoxygenate Mycin, 4-hydroxyperoxycyclophosphamide, 5-fluorouracil (5-FU), 5-fluorodeoxyuridine (5-FUdR), methotrexate (MTX), colchicine, paclitaxel, vincristine , vinblastine, etoposide (VP-16), trimetrexate, irinotecan, topotecan, gemcitabine, teniposide, cisplatin, and diethylstilbestrol (DES). In general, see The Merck Manual of Diagnosis and Therapy, 15th Ed. 1987, pp. 1206-1228, Berkow et al., eds., Rahway, N.J. When used with the compounds of the present invention, such chemotherapeutic agents can be used alone (for example, 5-FU and oligonucleotide), sequentially (for example, 5-FU and oligonucleotide for a period of time, followed by MTX and oligonucleotides), or in combination with one or more other such chemotherapeutic agents (for example, 5-FU, MTX and oligonucleotides, or 5-FU, a radiotherapeutic agent and oligonucleotides). Anti-inflammatory drugs (including but not limited to NSAIDs and corticosteroids) and antiviral drugs (including but not limited to ribavirin, vidarabine, acyclovir, and ganciclovir) can also be combined to in the composition of the present invention. See generally The Merck Manual of Diagnosis and Therapy, 15th Ed., Berkow et al., eds., 1987, Rahway, N.J., pp. 2499-2506 and 46-49, respectively). Other non-dsRNA chemotherapeutic agents are also within the scope of the invention. Compounds of two or more combinations may be used together or sequentially.

Toxicity and therapeutic efficacy of such compounds can be determined by standard pharmaceutical procedures in bacterial cultures or experimental animals, for example, to determine the LD50 (the dose lethal to 50% of the population) and the ED50 (the dose effective in treating 50% of the population). The ratio between toxic and therapeutic effects is the therapeutic coefficient and it can be expressed as the ratio LD50/ED50. Compounds with high therapeutic coefficients are preferred.

Data obtained from cell culture assays and animal studies can be used in formulating a dosage for human use. The dosage of compositions of the invention lies generally within a range of circulating concentrations that lie within the ED50 with little or no toxicity. The dosage may vary depending upon the dosage form employed and the route of administration utilized. For compounds used in the methods of the invention, the therapeutically effective amount can be estimated initially from cell culture assays. A dose can be formulated in animal models to achieve a circulating plasma concentration range of the compound, or, where appropriate, of the polypeptide product of the sequence of interest (e.g., to achieve a reduced polypeptide concentration), including determination of the IC50 in cell culture (ie, the concentration at which the test compound achieves a half-maximal inhibition of symptoms). Such information can be used to more accurately determine useful doses in humans. Levels in plasma can be measured, for example, by high performance liquid chromatography.

In addition to the administration discussed above alone or in combination, the dsRNAs of the invention can be administered in combination with other agents known to be effective in the treatment of influenza. In any event, the physician can adjust the amount and timing of dsRNA administration based on results observed using standard measures of potency known in the art.

Treatment Methods and Routes of Delivery

Compositions comprising an iRNA agent (eg, an iRNA agent targeted to influenza virus) can be delivered locally to a site of action or systemically into a subject by a variety of routes. Exemplary routes include direct topical application to affected areas, such as the lungs and nasal passages, as well as intravenous, nasal, buccal and ocular delivery. A preferred mode for administering the iRNA agents of the invention is by direct administration to the lung and nasal passages as a liquid, aerosol or spray solution.

Generally, the delivery of the iRNA agent of the present invention is the process of completing the delivery of the iRNA agent to the infected site in the subject. A preferred means of accomplishing this delivery is by topical administration to the lung or nasal passages, eg, to respiratory tissues by inhalation, spray or intranasal administration, or systemically, eg, by parenteral administration.

Dosage forms for inhalation or parenteral administration are known to those skilled in the art. Such dosage forms may comprise sterile aqueous solutions which may also contain buffers, diluents and other suitable additives. For intravenous use, the total concentration of solutes should be controlled so as to preserve the isotonicity of the formulation.

The active compounds disclosed herein are preferably administered to the lungs or nasal passages of a subject by any suitable means. Administration of the active compounds can be accomplished by administering an aerosol suspension containing respirable particles of one or more active compounds which are inhaled by the subject. The active compound can be aerosolized in various forms such as, but not limited to, dry powder inhaler, metered dose inhalant, or liquid/liquid suspension. Respirable particles can be liquid or solid. As described in U.S. Patent No. 4,501,729, the particles may optionally contain, with selected compounds such as amiloride, benzyl amiloride, or phenylamloride, effective in inhibiting passage of mucus from passageways. Other therapeutic ingredients in the amount of reabsorbed water in the secretions.

Granular pharmaceutical compositions can optionally be associated with carriers to aid in dispersion or transport. A suitable carrier such as sugar (eg glucose, lactose, sucrose, trehalose, mannitol) may be mixed with one or more active compounds in any suitable ratio (eg 1:1 weight ratio).

Particles containing the active ingredient used in the practice of this invention should include particles of respirable size, that is, particles of a size small enough to be inhaled through the mouth or nose and throat and to enter the bronchi and alveoli of the lungs. It is generally preferred that the particles have a size of about 1 to 10 microns (more particularly, sizes less than 5 microns). Aerosols contain particles of non-respirable size that tend to settle in the throat and be swallowed, so it is preferable to minimize the number of non-respirable particles in the aerosol. For nasal administration, particle sizes in the range of 10-500 microns are preferred to ensure their presence in the nasal cavity.

Liquid pharmaceutical compositions of the active compounds for the manufacture of aerosols can be prepared by combining the active compounds with a suitable excipient, such as sterile pyrogen-free water. The hypertonic saline solution used in the practice of the present invention is preferably a sterile pyrogen-free solution containing 1-15% by weight of physiologically acceptable salts, and preferably 3-7% by weight of physiologically acceptable salts.

Aerosols of liquid particles containing the active compound may be prepared by any suitable means, for example by use of a pressure-driven jet nebulizer or an ultrasonic nebulizer. See, for example, U.S. Patent No. 4,501,729 on this subject. Nebulizers are commercially available devices that convert a solution or suspension of an active ingredient into a pharmaceutical aerosol spray by accelerating a compressed gas (usually air or oxygen) through a small orifice or by ultrasonic agitation .

A suitable dosage formulation for use in a nebuliser consists of a liquid carrier and an active ingredient in an amount of up to 40% w/w of the dosage formulation, but preferably less than 20% w/w. The vehicle is usually water (most preferably sterile pyrogen-free water) or a diluted aqueous alcoholic solution, preferably an isotonic vehicle, but can be rendered hypertonic with respect to body fluids by the addition of, for example, sodium chloride. Optional additives include preservatives such as methylparaben (used when the dosage form formulation is not made sterile), antioxidants, flavoring agents, volatile oils, buffers and surfactants.

Aerosols of solid particles containing the active ingredient can be similarly produced using any solid particle therapeutic agent aerosol generator. An aerosol generator for administering a solid particulate therapeutic agent to a subject generates respirable particles at a rate suitable for human administration and generates a volume of aerosol containing a predetermined dose of the therapeutic agent. One exemplary type of solid particle aerosol generator is an insufflator. Suitable dosage formulations for administration by insufflation include finely divided powders, which may be delivered by means of an insufflator, or which may be absorbed into the nasal cavity by means of snuff. In an insufflator, a powder (e.g., a predetermined dose of powder effective to carry out the treatments described herein) is contained in a capsule or cartridge, usually made of gelatin or plastic, placed in the It is pierced or opened in situ, inhaled to allow the powder to be air drawn through the device, and delivered, or delivered by a manually operated pump. Powders for filling the inhaler may consist of the active ingredient alone or of a powder mix containing the active ingredient, a suitable powder diluent such as lactose, and optionally a surfactant. Dosage formulations usually contain 0.1 to 100 w/w active ingredients.

A second type of exemplary aerosol generator comprises a dose inhaler. Dosage inhalers are pressurized aerosol dispensers, usually containing a suspension or solution of the active ingredient in a liquefied propellant. With these devices, the formulation may be released through a valve so that the formulation is delivered in a given volume (typically 10 to 200ul) to prepare a particle spray containing the active ingredient. Suitable propellers include certain chlorofluorocarbon compounds such as dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, mixtures thereof, and the like. The formulations may further contain one or more co-solvents (eg, ethanol), surfactants (eg, oleic acid or sorbitan trioleate), antioxidants, and suitable flavoring agents.

The iRNA agent can be added to a pharmaceutical composition suitable for administration. For example, a composition can include one or more iRNA agents and a pharmaceutically acceptable carrier. As used herein, "pharmaceutically acceptable carrier" is meant to include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isobaric absorption delaying agents, and the like, compatible with pharmaceutical administration. The use of such media and agents for pharmaceutically active substances is known to those skilled in the art. Its use in the compositions is contemplated in addition to any conventional media or agents incompatible with the active compounds. Supplementary active compounds can also be incorporated into the compositions.

Administration can be by the subject or another person such as a caregiver. A caregiver may be any subject who provides care or care to a person. For example, a hospital, hospice, doctor's office, outpatient clinic; a healthcare worker such as a doctor, nurse, or other medical practitioner; or a guardian such as a spouse or parent. The drug therapy may be provided in metered doses, or with a dispenser that delivers the prescribed dose.

The term "therapeutically effective amount" refers to an amount of a composition present to provide the desired level of the composition of drug in the subject to be treated to obtain the desired physiological response.

The term "physiologically effective amount" refers to an amount that is delivered to a subject to produce the desired palliative or therapeutic effect.

The term "pharmaceutically acceptable carrier" means that the carrier can be absorbed into the lungs without causing significant toxic side effects to the lungs.

The term "co-administration" refers to the administration of the two or more agents, especially two or more iRNA agents, into a subject. The agents may be contained in a single pharmaceutical composition and administered simultaneously, or the agents may be contained in separate dosage formulations and administered sequentially to the subject. Two agents are said to be administered simultaneously as long as the two agents are simultaneously detectable in the subject.

Types of pharmaceutical excipients that can be used as carriers include: stabilizers such as human serum albumin (HSA); bulking agents such as carbohydrates, amino acids, and peptides; pH adjusters or buffers; salts such as sodium chloride ;etc. These supports may be crystalline or amorphous, or a mixture of the two.

Particularly valuable fillers include compatible carbohydrates, polypeptides, amino acids or combinations thereof. Suitable carbohydrates include: monosaccharides such as galactose, D-mannose, and sorbose; disaccharides such as lactose and trehalose; cyclodextrins such as 2-hydroxypropyl-β-cyclodextrin; raffinose, maltodextrin Polysaccharides such as essence and dextran; alditols such as mannitol and xylitol; and so on. Included in a preferred group of carbohydrates are lactose, trehalose, raffinose, maltodextrin and mannitol. Suitable polypeptides include aspartame. Amino acids include alanine and glycine, preferably glycine.

Suitable pH adjusters or buffers include, for example, sodium citrate, sodium ascorbate, and the like, organic salts prepared with organic acids and bases, preferably sodium citrate.

dose

The iRNA agent can be administered at a unit dose of less than about 75 mg/kg body weight, or at less than about 70, 60, 50, 40, 30, 20, 10, 5, 2, 1, 0.5, 0.1, 0.05, 0.01, 0.005, 0.001 or 0.0005 mg/kg body weight, and less than 200 nmol of iRNA agent (e.g., about 4.4 x ¹⁰ copies (copies))/kg body weight or less than 1500, 750, 300, 150, 75, Doses of 15, 7.5, 1.5, 0.75, 0.15, 0.075, 0.015, 0.0075, 0.0015, 0.00075, 0.00015 nmol of iRNA agent/kg body weight were administered. The unit dose may be administered, for example, by injection (eg, intravenously or intramuscularly, intrathecally or directly into an organ), inhalation or topically.

The dosage of the iRNA agent administered directly to an organ (e.g., to the lung) is about 0.00001 mg to about 3 mg/organ, or preferably about 0.0001-0.001 mg/organ, about 0.03-3.0 mg/organ, about 0.1-3.0 mg/eye or About 0.3-3.0mg/organ.

The dosage may be an amount effective for the treatment or prevention of the disease or disorder. It can be given as prophylaxis or as a main or part of a prescription.

In one embodiment, the unit dosage may be less than once per day, eg less than every 2, 4, 8 or 30 days. In other embodiments, the unit dosage is not administered on a frequent basis (eg, on a regular basis). For example, a unit dose may be administered in a single dose. Since iRNA agent-mediated silencing can persist for several days following administration of an iRNA agent composition, in many cases the composition can be administered less frequently than daily, or, in some cases, can be administered Administer only once throughout the treatment regimen.

In one embodiment, the subject is administered an initial dose and one or more maintenance doses of an iRNA agent, such as a double-stranded iRNA agent or an siRNA agent (e.g., such as an iRNA agent capable of being processed into an siRNA agent precursors such as larger iRNA agents, or DNA capable of encoding iRNA agents such as double-stranded iRNA agents or siRNA agents, or precursors thereof, etc.). The maintenance dose(s) are usually lower than the starting dose, eg, less than half the starting dose. The maintenance mode may include treating the subject with one or more doses within the dose range of 0.01-75 mg/kg body weight/day, such doses may be 70, 60, 50, 40, 30, 20, 10, 5, 2, 1, 0.5, 0.1, 0.05, 0.01, 0.005, 0.001 or 0.0005 mg/kg body weight/day. The maintenance dose is preferably administered no more frequently than once every 5, 10, or 30 days. Further, the treatment regimen can be continued for a period of time that depends on the specific disease characteristics, the severity of the disease, and the overall condition of the patient. In preferred embodiments, the dosage may be delivered no more frequently than once a day (eg, no more than every 24, 36, 48 or more hours, such as no more than every 5 or 8 days). Following treatment, one can be monitored for changes in condition and for relief of disease symptoms. The dose of the compound may be increased when the patient has no apparent response to the current dose level; or the dose should be decreased when remission of disease symptoms has been observed, or when the disease state has disappeared, or when undesired side effects are observed .

The effective amount can be administered alone or in two or more doses as needed or considered appropriate in specific cases. If desired to facilitate repeated or frequent infusions, implant delivery devices, for example, pumps, semi-permanent stents (e.g., intravenous, intraperitoneal, intracisternal or intracapsular), or reservoirs ( reservoir).

Following successful treatment, maintenance therapy is required in patients to prevent recurrence of the disorder, wherein the compound of the invention is administered at a maintenance dose of 0.001 to 100 g/kg body weight (see US 6,107,094).

The concentration of the iRNA agent composition is an amount effective enough to treat or prevent a disorder or modulate a physiological condition in a human. The concentration or amount of iRNA agent administered depends on the parameters established for the agent and the method of administration (eg, nasal, buccal, or pulmonary). For example, nasal administration tends to require lower concentrations of some ingredients to avoid irritation or burning of the nasal passages. Sometimes it is necessary to dilute the oral dosage form 10 to 100 times to provide a suitable nasal dosage form.

Certain factors may affect the dosage effective to treat a subject, including, but not limited to, the severity of the disease or condition, previous treatments, the general health and/or age of the subject, and any other diseases etc. It is also preferred if the effective dose of an iRNA agent, such as an siRNA agent, used for therapy can be increased or decreased over the course of a particular treatment. Variations in dosage may be due to, and are evident from, the results of diagnostic assays. For example, the subject can be monitored after administration of the iRNA agent composition. Additional amounts of the iRNA agent composition can be administered based on the information obtained from the monitoring.

Dosage depends on the severity and responsiveness of the condition being treated, over several days to several months of treatment, or until cure or remission is achieved. Optimal dosing schedules can be calculated from measurements of drug accumulation in the patient's body. Optimum dosages, dosing methods and repetition rates are readily determined by those skilled in the art. Optimum dosages may vary depending on the relative potency of each compound and can be estimated based on the above EC50s effective in in vitro and in vivo animal models.

The present invention will be described below using examples, but the examples should not be construed as further limiting the present invention.

Example

The nucleic acid sequences are represented below using standard nomenclature and the abbreviations specified in Table 3.

Table 3 Abbreviations for Nucleotide Monomers Used in Nucleic Acid Sequence Representations. It should be understood that these monomers are linked to each other by 5'-3'-phosphodiester linkages in the oligonucleotide.

abbreviation ^a Nucleotide A, a 2'-Deoxy-Adenosine-5'-Phosphate, Adenosine-5'-Phosphate C,c 2'-Deoxy-cytidine-5'-phosphate, Cytidine-5'-phosphate G, g 2'-Deoxy-guanosine-5'-phosphate, Guanosine-5'-phosphate T, t 2'-Deoxy-thymidine-5'-phosphate, Thymidine-5'-phosphate U, u 2'-Deoxy-uridine-5'-phosphate, Uridine-5'-phosphate N, n Any 2'-deoxy-nucleotide/nucleotide (G, A, C, or T, g, a, c, or u) am 2'-O-methyladenosine-5'-phosphate cm 2'-O-methylcytidine-5'-phosphate gm 2'-O-methylguanosine-5'-phosphate tm 2'-O-methyl-thymidine-5'-phosphate um 2'-O-methyluridine-5'-phosphate A , C , G , T , U , a , c , g , t , u Underlined: nucleoside-5'-phosphorothioate x universal base

^a Capital letters represent 2'-deoxyribonucleotides (DNA), lowercase letters represent ribonucleotides (RNA)

source of reagents

Where the source of reagents is not specified, such reagents may be obtained from any supplier of molecular biology reagents of a quality/purity standard for molecular biology applications.

Example 1: Selection sequence

siRNA design was performed to determine the influenza A mRNAs of the MP, NP, PA, PB1 and PB2 proteins targeted by the siRNAs. In the first round, insilico selection of siRNAs had 44 sequences targeting MP, 3 sequences targeting NP, and 1 sequence targeting PB1. There are no siRNAs specific for influenza A genes PA or PB2, 80% target coverage and 80% target efficiency are required by the first step selection process (see below).

 9.3操作系统下的工作站上。为了运行perl脚本(perl scripts)，确保在标准设置的Suse Linux 9.3下perl编译器(perl interpreter)(版本5.8.6(copyright 1987-2004，Larry Wall))能工作。从North Carolina State University网站上的Brown Lab Web服务器(Brown Lab Web Server)上下载BioEdit序列比对编辑程序(Hall，T.A.，Nucl.Acids.Symp.Ser.1999，41：95)，并安装于Microsoft Windows2000

操作系统下的计算机上。In order to build an environment for sequence analysis, download the fastA software package (Pearson, WR, & Lipman, DJ, PNAS 1988, 85: 2444) from ftp://ftp.virginia.edu/pub/ , and install it in the Suse

On a workstation under the 9.3 operating system. To run perl scripts, make sure the perl interpreter (version 5.8.6 (copyright 1987-2004, Larry Wall)) works under the standard setup of Suse Linux 9.3. Download the BioEdit sequence alignment editing program (Hall, TA, Nucl.Acids.Symp.Ser.1999, 41:95) from the Brown Lab Web server (Brown Lab Web Server) on the North Carolina State University website, and install it on Microsoft Windows2000

on the computer under the operating system.

The electronic selection workflow is as follows: download the required influenza A sequences, perform alignments, and generate statistics to obtain the distribution of bases at each position relative to the calculated consensus sequence. Candidate target regions meeting defined cut-off criteria were identified using a perl script. The specificity of the siRNA sequences of the candidate target regions to the human RefSeq database was analyzed using the fastA algorithm. siRNAs were scored according to predicted specificity with another perl script. Finally, siRNAs that meet specificity criteria are manually selected.

The required influenza A sequence was downloaded from the NCBI influenza virus database on June 24, 2005, and the download website is the website of National Center for Biotechnology Information. The number of each gene for MP, NP, PB1, PB2, and PA is shown in Table 4, and the corresponding accession numbers are given in Table 4.

Table 4: Number of gene sequences for influenza genes MP, NP, PB1, PB2 and PA from various virus subtypes used in siRNA sequence electronic selection

Genes H1N1 H2N2 H3N2 H5N1 H7N3 H7N7 H9N2 Total

MP 10 2 13 166 28 16 128 363

NP 12 3 10 169 12 6 138 350

PB1 3 2 10 10 163 10 7 127 322

PB2 2 1 10 164 12 9 133 331

PA 2 2 10 171 11 8 124 328

A global alignment of all sequences was generated using the ClustalW multiple alignment function of the BioEdit sequence alignment editing program (Thompson, J.D., et al., Nucleic Acids Res. 1994; 22:4673), using default parameters for each target separately. A positional nucleotide numerical summary output is obtained, providing information on the distribution of bases at each position relative to the calculated consensus sequence for each target.

The cut-off criteria for identifying candidate targeting regions of 19 nucleotides in length were defined as:

Criterion 1, target coverage: at least 80% of all sequences available for each influenza A gene need to be represented in the candidate region.

Criterion 2, targeting efficiency: at least 80% of all sequences - where candidate regions are represented, need to be identical in said candidate regions.

Criterion 1 was defined in order to avoid no sequence information in the region. Criterion 2 ensures that a large number of isoforms are targeted.

Nucleotide numerical profiles of positions were screened with a perl script to identify candidate target regions meeting the cleavage criteria with a length of 19 bases and to generate files that were used as fastA input in subsequent analysis steps. For script input, enter the total number of sequences for each target and enter a value of 80 for the percent conserved. All candidate sense siRNAs corresponding to the most common sequences in the candidate target region were selected and saved in a fastA format file. To account for interactions between potential dTdT overhangs of siRNAs and target sequences, all sequences were extended with "AA" at the 5' end, resulting in a 21-mer input sequence. For each candidate target region, another file is generated with the region characteristics: target coverage (sequences present), targeting efficiency, total number of mismatches, number of conserved sequences, and sequences present number.

For further selection, candidate siRNAs were ranked according to their predicted potential to interact with host (here, without limitation, human) genes (off-target potential). It is assumed that siRNAs with low off-target potential are more specific in vivo.

To predict siRNA-specific off-target potential, the following assumptions were made:

1) Positions 2–9 of the strand (counting from 5' to 3') contribute more to off-target potential than the rest of the sequence (non-seed region) (Haley, B., and Zamore, P.D., Nat Struct Mol Biol. 2004, 11:599).

2) An off-target score is calculated for each hit, based on the identity to the siRNA sequence and the position of the mismatch.

3) The sense strand can be made RNA interference resistant by introducing appropriate nucleotide modifications into the sense strand (eg, all nucleotides containing pyrimidine bases are 2'-O-methyl modified nucleotides) Inactive; therefore, only the off-target potential of the antisense strand needs to be considered.

To identify potential off-target genes, a homology search was performed against publicly available human mRNA sequences on 21mer sequences corresponding to candidate target regions plus a 3' terminal AA tail (to account for TT overhangs). To this end, a fastA (version 3.4) search was performed against the human RefSeq database (available version downloaded from ftp://ftp.ncbi.nih.gov/refseq/ on July 27, 2005) with all 21mer input sequences. A fastA search was performed with parameters-values-pairs-b 30 -g 30, thereby calculating the homology of the 21 mer full-length sequence. The search results display the potential off-targets of the candidate siRNA sequences in tabular form.

To sort the resulting list of potential off-target data, the fastA output files were analyzed to identify the host genes with the highest off-target values. The following off-target properties were extracted for each 21mer input sequence, and an off-target score was calculated for each potential off-target.

1. The number of identical nucleotides in the 21mer sequence (identity)

2. The number of mismatches in the seed region

For hypotheses 1 and 2 considered, the off-target score was calculated as follows:

Identity - 0.2 ^* number of seed mismatches

All siRNAs were sorted according to their highest off-target score (rising). An off-target score of 16.8 was used as a cut-off point for siRNA selection. 42 siRNAs specific to influenza A matrix protein (MP), 3 siRNAs specific to influenza A nucleocapsid protein (NP) and 1 siRNA specific to influenza A polymerase basic protein 1 (PB1) Off-target scores at or below this threshold.

Given the relatively low number of candidate siRNAs generated by the selection method described above, the nuclei for the positions were rechecked with a cut-off point of criterion 1 (target coverage) set at 70% and criterion 2 (targeting efficiency) maintained at 80%. Numerical summary of nucleotides, followed by repeating the ranking of off-target scores as described above. For a total of 48 candidate siRNAs, two additional siRNAs specific to influenza A MP mRNA were selected with a targeting efficiency of 79.9%. The sequences of these 48 candidate siRNAs are shown in Table 1A.

Because the selection methods described above yielded only a limited number of candidate agents, the selection criteria were slightly relaxed to generate additional candidate agents. Specifically, criterion 1 above was relaxed to 50% target coverage, while criterion 2, targeting efficiency, was kept at 80%, and the selection process described above was repeated. This approach yielded additional reagents AL-DP-8001 to AL-DP-8040, which are listed in Table 1C.

In such a process, it can be appreciated that the off-target scoring step results in the greatest attrition rate in potential reagents. In order to obtain more candidate agents, the selection process was repeated one more time with criterion 1 for 80% target coverage and criterion 2 for 80% targeting efficiency, and off-target scores were ignored. Additional formulations obtained through this process are listed in Table 1D. But the other candidate agents listed in Table 1E were obtained by repeating this selection again with criterion 1 for 50% target coverage, criterion 2 for 80% targeting efficiency and ignoring the off-target score.

Additional candidate iRNA agents were identified by allowing the introduction of free bases. When introducing a maximum of 3 free bases per strand in the non-seed region of the iRNA reagent (corresponding to positions 2–9 of the antisense strand), first identify candidates with 100% target coverage and targeting efficiency using a perl script sequence. Table IF shows reagents identified in this way. In the second round, additional iRNAs were identified that had 80% target coverage and targeting efficiency when allowing the introduction of 1 free base. These iRNAs are listed in Table 1G.

Example 2: siRNA synthesis

Synthesis of Nucleotides Containing Natural Bases

，Glen Research，Sterling VA)作为固体支持物。RNA和含有2’-O-甲基核苷酸的RNA分别使用相应的亚磷酰胺和2’-O-甲基亚磷酰胺通过固相合成产生(Proligo Biochemie GmbH，Hamburg，Germany)。将这些结构单元(building blocks)引入到寡核苷酸链序列范围内的所选择的位点上，其利用标准核苷亚磷酰胺化学，如在书《Current protocolsin nucleic acid chemistry》，Beaucage，S.L.等(Edrs.)，John Wiley & Sons，Inc.，New York，NY，USA中所描述。硫代磷酸键通过将碘氧化剂溶液替代为在乙腈(1％)中的Beaucage试剂(Chruachem Ltd，Glasgow，UK)溶液而引入。进一步的辅助试剂获自Mallinckrodt Baker(Griesheim，Germany)。Single-stranded RNAs were solid-phase synthesized at a scale of 1 μmole using an Expedite 8909 synthesizer (Applied Biosystems, AppleraDeutschland GmbH, Darmstadt, Germany), controlled pore glass (CPG, 500

, Glen Research, Sterling VA) as a solid support. RNA and RNA containing 2'-O-methyl nucleotides were produced by solid phase synthesis (Proligo Biochemie GmbH, Hamburg, Germany) using the corresponding phosphoramidites and 2'-O-methylphosphoramidites, respectively. These building blocks are introduced at selected sites within the sequence of the oligonucleotide chain using standard nucleoside phosphoramidite chemistry, as described in the book "Current protocols in nucleic acid chemistry", Beaucage, SL (Edrs.), John Wiley & Sons, Inc., New York, NY, USA. Phosphorothioate linkages were introduced by replacing the iodine oxidizing agent solution with a solution of Beaucage reagent (Chruachem Ltd, Glasgow, UK) in acetonitrile (1%). Further auxiliary reagents were obtained from Mallinckrodt Baker (Griesheim, Germany).

Deprotection and purification were performed according to existing methods by anion-exchange HPLC on crude oligonucleotides. The yield and concentration were determined by the UV absorbance of each RNA solution at a wavelength of 260 nm using a spectrophotometer (DU640B, Beckman Coulter GmbH, Unterschleiβheim, Germany). Double-stranded RNA was formed by mixing with an equimolar complementary strand solution in annealing buffer (20mM sodium phosphate, pH 6.8; 100mM sodium chloride), heated in a water bath at 85-90°C for 3 minutes, and then passed through 3- 4 hours to cool to room temperature. The purified RNA solution was stored at -20°C until use.

As a result of the synthesis procedure described above, all oligonucleotides synthesized as described above do not have a phosphate group on the 5'-terminal nucleotide.

Synthesis of nucleotides containing universal bases

5'-O-(4,4'-dimethoxytrityl)-2'-O-(tert-butyldimethylsilyl)-1'-(5-nitroindolyl)- Synthesis of Phosphoramidites of D-riboside and Controlled Pore Glass Support

Step A: 1-O-methyl-D-riboside (102).

To a solution of D-ribose (25 g) in dry methanol (300 mL) was added concentrated sulfuric acid (1.88 mL), and stirred at room temperature for 3 days. The reaction mixture was then neutralized with 1N sodium hydroxide solution and concentrated to a crude residue. The crude residue was dissolved in methanol (200 mL), and the solid was filtered off. The filtrate was concentrated to a crude residue, then applied to a silica gel column and eluted with dichloromethane-methanol (5:1) to give pure compound (23.0 g, 82%) as syrup.

Step B: 1-O-methyl-2,3,5-tri-O-(2,4-dichlorobenzyl)-D-riboside (103)

To a solution of 1-O-methyl-D-riboside (13.43 g, 81.83 mmol), 18-crown-6 (1.34 g) in dry THF (100 mL) was added potassium hydroxide powder (69 g, 1.23 mol), And stirred at room temperature for 40-60min. 2,4-Dichlorobenzyl chloride (51 mL, 368.2 mmol) was added dropwise and the reaction mixture was stirred overnight at the same temperature. The solid was filtered off, and the filtrate was concentrated to a crude residue, which was then applied to a silica gel column and eluted with hexane-ethyl acetate (4:1) to give pure compound (48 g, 92%) as a white solid.

1H-NMR (CDCl ₃ , 400MHz): δ 7.46-7.34(m, 5H, ArH), 7.24-7.16(m, 4H, ArH), 4.99(s, 1H, H-1), 4.71(dd, 2H, Jgem=12.8Hz, OCH ₂ Ar), 4.63-4.61 (m, 4H, 2 OCH ₂ Ar), 4.38-4.36 (m, 1H), 4.19-4.16 (dd, 1H), 3.98 (d, 1H, J= 4.4Hz), 3.75(dd, 1H, J=3.6, J=10.2Hz, H-5a), 3.66(dd, 1H, J=3.6, J=10.4Hz, H-5b), 3.37(s, 3H, OCH ₃ ).

Step B: 1-Bromo-2,3,5-tri-O-(2,4-dichlorobenzyl)-D-ribose (104).

To 1-O-methyl-2,3,5-tri-O-(2,4-dichlorobenzyl)-D-riboside (3.22 g, 5.02 mmol) in dry dichloromethane cooled in an ice bath (50 mL) was added HOAc-HBr (5.3 mL, 30%) and stirred at 0-25 °C for 3 h. The reaction mixture was concentrated to a crude residue and co-evaporated with toluene (3 x 30 mL) to give the crude residue, which was dried under good vacuum and used in the next reaction without purification and identification as a syrup.

Step D: 1-(5-Nitroindolyl)-2,3,5-tri-O-(2,4-dichlorobenzyl)-D-riboside (105).

To a solution of 5-nitroindole (2.44 g, 15.06 mmol) in dry _CH3CN (30 mL) was added sodium hydride (602 mg, 15.06 mmol, 60%) and stirred at room temperature under argon for 3-4 h. The previously obtained sugar donor (104) in dry _CH3CN (10 mL) was added and stirred overnight at the same temperature under argon. The solid was filtered off and the filtrate was concentrated to a crude residue which was applied to a silica gel column and eluted with hexane-ethyl acetate (3:1) to give pure compound 105 (2.16 g, 60%) as α and β Mixture (1:1).

Step E, F: 5'-O-(4,4'-dimethoxytrityl)-1'-(5-nitroindolyl)-D-riboside (106) and (107)

To 1-(5-nitroindolyl)-2,3,5-tri-O-(2,4-dichlorobenzyl)-D-riboside 105 (1.16g, 1.51mmol) at -78°C A cold solution of dry dichloromethane (100 mL) was added to a solution of _BCl3 in dichloromethane (23 mL, 1.0 M) and stirred under argon at the same temperature for 2 h and at -40 °C for 2 h. The reaction mixture was cooled with methanol-dichloromethane (1:1, 50 mL) and neutralized with ammonia-methanol solution. The solid was filtered off, and the filtrate was concentrated to a crude residue, which was applied to a silica gel column and eluted with dichloromethane-methanol (10:1) to give the pure compound (300 mg, 68%) as an alpha and beta mixture (1 :1). To a solution of the compound obtained above (840 mg, 2.86 mmol) in dry pyridine (3-4 ml) and DMAP (90 mg) was added DMTrCl (1.06 g) and stirred overnight at room temperature under argon. The reaction mixture was concentrated to a crude residue which was added to a silica gel column and eluted with hexane-ethyl acetate (1:1) to give pure compound 106 (550 mg) and compound 107 (190 mg), a mixture of compounds 106 and 107 (360mg).

Compound 106: 1H-NMR (CDCl ₃ , 2D g-COSY and 2D NOESY, 400MHz): δ8.49(d, 1H, J=1.6Hz), 8.35(d, 1H), 8.03(dd, 1H, J= 2.0, J=9.0Hz), 7.70-7.69(m, 2H), 7.47-7.14(m, 8H, ArH), 6.86-6.81(m, 5H, ArH), 6.71(d, 1H, J=3.6Hz) , 6.41(d, J=5.2Hz, H'-1), 4.73(t, 1H, J=4.8Hz, H'-2), 4.46-4.42(m, 3H, H'-3, H'-4 , H'-5), 3.79 (s, 6H, 2OCH ₃ ), 3.51 (dd, 1H, J=3.2, J=10.4Hz, H'-5a), 3.26 (dd, 1H, J=3.2, J= 10.6Hz, H'-5b.

Compound 107: 1H-NMR (CDCl ₃ , 2Dg-COSY and 2D NOESY, 400MHz): δ8.55 (d, 1H, J=2.0Hz), 7.98 (dd, 1H, J=2.4, J=9.2Hz), 7.60(d, 1H, J=9.2Hz), 7.53(d, 1H, J=3.2Hz), 7.44-7.42(m, 2H), 7.34-7.24(m, 7H, ArH), 6.84-6.81(m, 4H, ArH), 6.68(d, 1H, J=3.2Hz), 6.00(d, 1H, J=5.2Hz, H'-1), 4.53(t, 1H, J=7.6Hz), 4.46-4.44( m, 1H), 4.23-4.20(m, 1H), 3.80-3.76(m, 7H, 2OCH ₃ , H'-5), 3.55(dd, 1H, H'-5a), 3.43(dd, 1H, H '-5b).

Step G: 5'-O-(4,4'-dimethoxytrityl)-2'-O-(tert-butyldimethylsilyl)-1'-(5-nitroindole base)-D-riboside (108) and 5'-O-(4,4'-dimethoxytrityl)-3'-O-(tert-butyldimethylsilyl)-1' -(5-nitroindolyl)-D-riboside (109)

To 5'-O-(4,4'-dimethoxytrityl)-1'-(5-nitroindole)-D-riboside (106) (550mg, 0.92mmol), AgNO ₃ ( 188 mg, 1.104 mmol), and pyrimidine (0.74 mL, 9.2 mmol) in dry THF (9.2 mL) was added TBDMSCl (188 mg, 1.196 mmol) and stirred overnight at room temperature under argon. The solid was filtered off, and the filtrate was concentrated to a crude residue, which was applied to a silica gel column and eluted with hexane-ethyl acetate (4:1) to give pure compound 108 (230 mg, 35%), compound 109 (150 mg, 23%), and the mixture of compounds 108 and 109 (110 mg, 17%), the total yield was 75%.

Compound 108: 1H-NMR (CDCl ₃ , 2D g-COSY, 2D NOESY, 400MHz): δ 8.56 (d, 1H, J=2.4Hz), 7.88 (dd, 1H, J=2.4, J=8.8Hz), 7.62(d, 1H, J=9.2Hz), 7.54(d, 1H, J=3.6Hz), 7.46-7.44(m, 2H), 7.36-7.25(m, 6H, ArH), 6.85-6.83(d, 5H, ArH), 6.69 (d, 1H, J=3.6Hz), 5.94 (d, 1H, J=7.2Hz, H'-1), 4.69 (dd, 1H, H'-2), 4.31-4.29 ( m, 2H, H'-3, H'-4), 3.80(s, 6H, 2OCH ₃ ), 3.58(dd, 1H, J=2.0, J=10.6Hz, H'-5a), 3.40(dd, 1H, J=2.0, J=10.4Hz, H'-5b), 2.85(d, 1H, J=0.8Hz, 3'-OH), 0.78(s, 9H, t-Bu), -.016(s , 3H, SiCH ₃ ), -0.43 (s, 3H, SiCH ₃ ).

Compound 109: 1H-NMR (CDCl ₃ , 2D g-COSY, 2D NOESY, 400MHz): δ 8.61 (d, 1H, J=2.4Hz), 8.05 (dd, 1H, J=2.0, J=8.8Hz), 7.69-7.65(m, 2H), 7.47-7.45(m, 2H, ArH), 7.36-7.27(m, 5H, ArH), 6.86-6.83(m, 3H, ArH), 6.71(d, 1H, J= 3.2Hz), 5.99(d, 1H, J=4.8Hz, H'-1), 4.51(t, 1H, J=4.8Hz, J=5.6Hz, H'-3), 4.40-4.36(m, 1H , H'-2), 4.17-4.15 (m, 2H, H'-4, H'-5), 3.82 (s, 3H, OCH ₃ ), 3.81 (s, 3H, OCH ₃ ), 3.63 (dd, 1H, J=2.4, J=11.0Hz, H'-5a), 3.31(dd, 1H, J=2.8, J=11.0Hz, H'-5b), 2.95(d, 1H, J=6.0Hz, 2 '-OH), 0.91 (s, 9H, t-Bu), 0.05 (s, 3H, SiCH ₃ ), 0.00 (s, 3H, SiCH ₃ ).

Step H: 5'-O-(4,4'-dimethoxytrityl)-2'-O-(tert-butyldimethylsilyl)-1'-(5-nitroindole base)-D-riboside-3'-O-cyanoethyl-N, N-diisopropyl phosphoramidate (110)

2-Cyanoethyl-N,N-diisopropylphosphoramidite chloride (153 mg, 0.646 mmol) was added to 5'-O-(4,4'-dimethoxytrityl)-3'- O-(tert-butyldimethylsilyl)-1-(5-nitroindolyl)-D-β-riboside 108 (230mg, 0.323mmol), diisopropylethylamine (306uL, 1.78 mmol) and DMAP (10 mg) in dry dichloromethane (3 mL) was stirred at room temperature under argon for 4-6 h. The reaction mixture was concentrated to a crude residue which was applied to a silica gel column saturated with 2% triethylamine in hexane and eluted with hexane-ethyl acetate (2:1) to afford pure compound as an amorphous solid 110 (250mg, 85%.

³¹ P-NMR (CDCl3, 400MHz): δ 149.54(s), 146.57(s). Anal. Cald of C ₅₀ H ₆₅ N ₄ O ₉ PSi: 924.43. Found: 947.43 [M+Na]+.

Step I: 2'-hydroxyl or 3' of 5'-O-(4,4'-dimethoxytrityl)-1-(5-nitroindolyl)-D-riboside (111) -Hydroxyl solid support.

Addition of succinic anhydride to 2'-OTBDMS of 5'-O-(4,4'-dimethoxytrityl)-1-(5-nitroindolyl)-D-β-riboside (109) (108) or a mixture of 3'-O-TBDMS and DMAP in solution in dry dichloromethane. The reaction mixture was stirred at room temperature under argon for 6 h. Succinic anhydride and another portion of DMAP were added and stirred for a total of 16h. The mixture was concentrated to a crude residue which was dissolved in ethyl acetate (50ml), washed with citric acid ( _400mg /20ml), brine and dried ( _Na2SO4 ). The organic layer was concentrated to crude nucleoside succinate. It was used directly in subsequent reactions without purification.

Succinic acid nucleoside, DMAP, DTNP and Ph ₃ P were stirred at room temperature for 20 min [Nucleoside and nucleotides, 1996, 15(4), 879-888.]. Then lcaa-CPG was added and stirred at the same temperature for 45 min. The solid was filtered off and washed with _CH3CN , dichloromethane and ether. The solid support is dried, capped and washed under standard procedures to yield a solid support.

The resulting controlled glass support comprising nitroindole groups and phosphoramidites were used in the standard oligonucleotide synthesis described above for oligonucleotides comprising natural bases.

Example 3: In vitro testing of siRNA

The ability of iRNA agents to inhibit influenza virus replication was tested in vitro in human cell lines or in mice. The iRNA reagent is transfected into the cells, eg, by transfection or electroporation, allowed to act on the cells for a certain period of time (eg, 24 hours), and the level of infection determined by plaque formation or ELISA analysis. For several influenza genes expressed recombinantly in mammalian host cells, RNAi agents were tested for inhibition of target gene expression to complement these direct analyses.

Viruses and Cell Lines

Influenza virus A/PR/8/34 (PR8), subtype H1N1, was obtained from Charles River Laboratories (ATCC # VR-1469). A/WSN/33 (WSN), subtype H1N1, by Thomas Chambers, University of Kentucky, Lexington, KY. USA (see Castrucci, M.R., et al., J Virol. 1992, 66:4647), or Dr. Peter Palese , Mount Sinai School of Medicine New York City, NY, USA (see WO 04/028471). Virus stocks were propagated in the allantoic cavity of eggs with embryos at 34°C for 48-72h (PR8) or 37°C for 24h (WSN) (Tompkins, S.M., et al. Proc.Natl.Acad.Sci.2004, 101: 8682).

MDCK cells were obtained from the American Type Culture Collection (ATCC, Rockville MD, USA; ATCC # CCL-34) and cultured in 8% heat-inactivated fetal bovine serum (FBS), 2 mM L-glucose Aminoamide, 1 mM sodium pyruvate, 1.5 g/L sodium bicarbonate, and non-essential amino acids were grown in MEM at 37°C under a 5% _CO2 /95% air atmosphere.

Vero E6 green monkey kidney epithelial cells were obtained from ATCC (Rockville MD, USA, ATCC #CRL-1586) and treated with 4.5g/l D-glucose, 2mM L-glutamine, 110mg/l sodium pyruvate, Grow in DMEM supplemented with 10% fetal bovine serum (Hyclone, Cat # 30070.03) and 0.1% penicillin/streptomycin at 37°C in a 5% _CO2 /95% air atmosphere.

Cos-7 Vero cells were obtained from the German Collection of Microorganisms and Cell Cultures (DSMZ, Braunschweig, Germany, DSMZ # ACC 60) and grown in Dulbecco's MEM, 10% fetal bovine serum , 2mM L-glutamine, 1.2μg/ml sodium bicarbonate, 100u penicillin/100μg/ml streptomycin (Biochrom AG, Berlin, Germany).

Example 3.1: Detection of Plaque Formation

Cell culture, siRNA transfection and virus infection.

The day before transfection, MDCK cells were seeded in 24-well plates at 7.5 x ¹⁰⁴ cells/well in 0.5 ml of growth medium. On the day of siRNA transfection, MDCK cells were 80% confluent. Before transfection, 0.25 ml of growth medium was added to the cells.

Before adding cells, mix 1.5ml (50μl/well) Optimem I (Invitrogen) and 90μl (3μl/well) Lipofectamin 2000 (Invitrogen), enough for transfection of a 24-well plate, in a 2ml Sarstedt tube and store at room temperature. Incubate for 10-15 minutes. Add the appropriate amount of siRNA dissolved in annealing buffer to the Optimem/lipofectamine 2000 mixture to obtain the desired final concentration, mix, and incubate for an additional 15-25 minutes at room temperature. Then, 50 μl of siRNA/reagent complex was added dropwise to each well as indicated by the assay design. Plates were then shaken slowly to ensure thorough mixing and incubated for 14 hours at 37°C in a 5% _CO2 /95% air atmosphere.

Then, slowly aspirate the transfection medium, wash the cells once with 0.25-0.5ml PBS, and add 100μl MEM solution of different concentrations of PR8 to each well, incubate at 37°C for 1-2 hours, then add 0.5ml overlay culture base (MEM, 20mM HEPES, 0.075% NaHCO ₃ , 2mM glutamine, 0.6% agarose, 0.5μg/ml TPCK-trypsin), and put the plate in the incubator at 37°C in 5% CO ₂ /95% Incubate under air atmosphere for 48 hours. Plates were then mounted and immunostained for viral plaques as described below.

Immunostaining and virus quantification

48 hours host-infection, cells fixed in neutral buffered 10% formalin for 45 minutes, wells rinsed with PBS. The wells were then blocked with permeation buffer (1 x PBS, 2% FBS, 0.5% saponin, 0.1% sodium azide) for 15 minutes at room temperature, and 125 μl containing 0.5 μg/ml mouse anti-influenza A biotin was added Solution of labeled antibody MAB8258B (Chemicon). After incubation at room temperature for 1 hr, the wells were washed twice with PBS to remove unbound and antibody, and 125 μl of 1 μg/ml horseradish peroxidase (HRP)-conjugated streptavidin (Vector Laboratories) in PBS was added. Add to each well, incubate the plate for 45 min and wash three times with PBS. 200 μl TMB substrate (Vector Laboratories #SK-4400)/well was added to each well. Then incubate for 5-10 minutes at room temperature in the dark, stop the colorimetric reaction with distilled water, drain the water, and dry the plate. Stained influenza plaques were counted with an inverted light microscope at 4X magnification. Plaque-forming activity was compared with cells transfected with Lipofectamin only (mock treatment) and expressed as [(plaque-forming activity in treated cells)/(plaque-forming activity in mock-treated cells)] x 100=% residual infection rate

Embodiment 3.2: ELISA detection:

One day before transfection, MDCK or Vero cells were seeded in 96-well plates at 10 ⁴ cells/well in 0.1 ml growth medium. On the day of siRNA transfection, cells were 80% confluent. Before transfection, 44 μl of growth medium was added to the cells.

Mix 1.08ml (9μl/well) Optimem I (Invitrogen) and 42μl (0.35μl/well) Lipofectamin 2000 (Invitrogen), enough for transfection in a 96-well plate, in a 2ml Sarstedt tube and incubate at room temperature for 10- 15 minutes. Add the appropriate amount of siRNA dissolved in annealing buffer to the Optimem/lipofectamine 2000 mixture to obtain the desired final concentration, mix, and incubate for an additional 15-25 minutes at room temperature. Then, 10 μl of siRNA/reagent complex was added dropwise to each well as indicated by the assay design. Plates were then shaken slowly to ensure thorough mixing and incubated for 14 hours at 37°C in a 5% CO2/95% air atmosphere.

Then, the cells were washed once with PBS, infected with 50 μl per well of MEM with PR8 influenza virus, and incubated for 1-2 hours. Then, the plate was washed once with PBS and 200 μl MEM with 0.25/0.5 μg/ml (MDCK/VERO) trypsin was added. Two days after infection, plates were fixed in 10% buffered formalin for 15 min. The cells were washed with PBS, blocked with blocking buffer for 15 min at room temperature, and 50 μl containing 0.5 μg/ml biotin-labeled anti-influenza A monoclonal antibody MAB8258B (Chemicon) was added to each well. After incubation for 1 hr at room temperature, the wells were washed twice with PBS, and 50 μl/well of blocking buffer solution containing 1 μg/ml AP-conjugated streptavidin (Vector Laboratories) was added. After incubation for 45 min and washing 3 times with PBS, 100 μl of pNPP substrate solution per well was added. Plates were developed at room temperature in the dark and read at 405 nm.

Example 3.3: Inhibition of recombinant expression of influenza target genes by siRNA

MP (SEQ ID NO: 1453), NP (SEQ ID NO: 1454), PA (SEQ ID NO: 1455), PB1 (SEQ ID NO: 1456) and PB2 (SEQ ID NO: 1456) were synthesized by GENEART (Regensburg, Germany). : 1457) consensus sequence (see Table 5), and clone it into the GENEART standard vector. MP and PA were subcloned into psiCheck-2 (Promega, Mannheim, Germany) via AsiSI and NotI (both from NEBn, Frankfurt, Germany) sites, NP, (PB1) and PB2 were subcloned via XhoI and NotI by This resulted in a construct with the influenza gene between the stop codon and the Renilla luciferase polyA signal. Correct cloning was confirmed by end-sequencing by GATC Biotech (Konstanz, Germany).

transfection

Cos-7 cells were seeded at 1.5 x ¹⁰⁴ cells/well on white 96-well plates with clear bottoms (Greiner Bio-One GmbH, Frickenhausen, Germany) in 75 μl of growth medium. Immediately after seeding the cells, 50 ng of plasmid/well was transfected with Lipofectamine2000 (Invitrogen) as described below for siRNAs, the plasmid was diluted with Opti-MEM to a final volume of 12.5 μl/well, and the entire plate was mastermixed.

Four hours after plasmid transfection, siRNA transfection was performed in quadruplicate. For each well, 0.5 μl of Lipofectamine2000 (Invitrogen GmbH, Karlsruhe, Germany) was mixed with 12 μl of Opti-MEM (Invitrogen) and incubated at room temperature for 15 minutes. To achieve an siRNA concentration of 50 nM in a 100 μl transfection volume, 1 μl of 5 μM siRNA was mixed with 11.5 μl Opti-MEM per well, combined with Lipofectamine2000-Opti-MEM and incubated again for 15 minutes at room temperature. During incubation, the growth medium was removed from the cells and replaced with 75 μl/well of fresh medium. The siRNA-Lipofectamine2000 complex was added completely to the cells and the cells were incubated for 24 hours at 37°C, 5% CO ₂ in a humidified incubator (Heraeus GmbH, Hanau, Germany).

Cells were harvested by removing the growth medium and adding 150 μl of a 1:1 mixture of medium and Dual-Glo luciferase substrate from the Dual-Glo Luciferase Assay System (Promega, Mannheim, Germany). Luciferase assays were performed according to the manufacturer's protocol for the Dual-Glo luciferase assay and fluorescence was measured in a Victor-Light 1420 Luminescence Counter (Perkin Elmer, Rodgau-Jügesheim, Germany). Values obtained with Renilla luciferase were normalized to each value obtained with firefly luciferase. Values obtained with siRNAs directed against influenza genes were normalized by setting the values obtained with non-specific siRNAs to 100% (for the neomycin resistance gene).

The screened effective siRNAs were further identified by dose-response curves. According to the above procedure, the dose-response curves were drawn at the following siRNA concentrations: 100 nM, 25 nM, 6.3 nM, 1.6 nM, 400 pM, 100 pM, 24 pM, 6 pM, 1.5 pM, 380 fM. _IC50 values were determined by parametrized curve fitting using the XLfit program.

Table 5: Influenza genes MP (SEQ ID NO: 1453), NP (SEQ ID NO: 1454), PA (SEQ ID NO: 1455), PB1 (SEQ ID NO: 1456) and PB2 ( The viral consensus sequence of SEQ ID NO: 1457)

MP: Obtained from gi|13383290|gb|AB049165|Influenza A virus (A/parakeet/Chiba/1/97(H9N2)) membrane ion channel, M1, M2 viral consensus sequence of matrix protein, complete的编码序列ATGAGTCTTC TAACCGAGGT CGAAACGTAC GTTCTCTCTA TCATCCCGTC AGGCCCCCTC 60AAAGCCGAGA TCGCGCAGAG ACTTGAAGAT GTCTTTGCAG AGAAGAACAC AGATCTCGAG 120GCTCTCATGG AATGGCTAAA GACAAGACCA ATCCTGTCAC CTCTGACTAA GGGGATTTTA 180GGGTTTGTGT TCACGCTCAC CGTGCCCAGT GAGCGAGGAC TGCAGCGTAG ACGCTTTGTC 240CAGAATGCCC TAAATGGGAA TGGAGACCCA AACAACATGG ACAGGGCAGT TAAACTATAC 300AAGAAGCTGA AGAGGGAAAT AACATTCCAT GGGGCTAAGG AAGTTGCACT CAGTTACTCT 360GCTGGTGCAC TTGCCAGTTG CATGGGTCTC ATATACAACC GGATGGGAAC AGTGACCACA 420GAAGTGGCTC TTGGCCTAGT GTGTGCCACT TGTGAGCAGA TTGCAGATTC ACAACATCGG 480TCCCACAGGC AGATGGCGAC TACCACCAAC CCACTAATCA GACATGAGAA CAGAATGGTG 540CTGGCCAGCA CTACAGCTAA GGCTATGGAG CAGATGGCTG GATCAAGTGA GCAGGCAGCG 600GAAGCCATGG AAGTCGCAAG TCAGGCTAGG CAGATGGTGC AGGCAATGGA 660ACTCATCCTA GCTCCAGTGC AGGTCTAAAA GATAATCTTC TTGAAAATTT GCAGGCCTAC 720CAGAAACGAA TGGGGGTGCA GATGCAGCGA TTCAAGTGAT CCTCTCGTTG TTGCAGCAAG 780TATCATTGGG ATCTTGCACT TGATATTGTG GATTCTTGAT CGTCTTTTCT TCAAATGCAT 840TTATCGTCGC CTTAAATACG GTTTGAAAAG AGGGCCTTCT ACGGAAGGAG TACCTGAGTC 900TATGAGGGAA GAGTATCGAC AGGAACAGCA GAGTGCTGTG GATGTTGACG ATGGTCATTT 960TGTCAACATA GAGCTGGAGT AA 982SEQ ID NO：1453 NP: Obtained from the viral consensus sequence of the nucleoprotein gene of H5N1 gi|14326108|AF370122|Influenza A virus (A/Goose/Guangdong/3/97 (H5N1)), complete coding sequence CTCACTGAGT GACATCAAAA TCATGGCGTC TCAAGGCACC AAACGATCTT ATGAACAGAT 60GGAAACTGGT GGAGAACGCC AGAATGCTAC TGAGATCAGA GCATCTGTTG GAAGAATGGT 120TGGTGGAGTT GGGAGGTTTT ATATACAGAT GTGCACTGAA CTCAAACTCA GCGACTATGA 180AGGAAGGCTG ATTCAGAACA GCATAACAAT AGAGAGAATG GTTGTCTCTG CATTTGATGA 240AAGGAGGAAC AAATACCTGG AAGAACATCC CAGTGCGGGG AAGGACCCAA AGAAAACTGG 300AGGTCCAATC TACCGAAGAA GAGACGGGAA ATGGGTGAGA GAGCTGATTC TGTATGACAA 360AGAGGAGATC AGGAGAATTT GGCGTCAAGC GAACAATGGA GAAGATGCAA CTGCTGGTCT 420CACTCACCTG ATGATCTGGC ATTCCAATCT AAATGATGCC ACATACCAGA GAACAAGAGC 480TCTCGTGCGT ACTGGGATGG ACCCTAGAAT GTGCTCTCTG ATGCAAGGAT CAACTCTCCC 540GAGGAGATCT GGAGCTGCTG GTGCGGCAGT AAAGGGAGTC GGAACTATGG TGATGGAACT 600AATTCGGATG ATAAAGCGAG GGATTAACGA TCGGAATTTC TGGAGAGGTG AAAATGGGCG 660 AAGAACAAGG ATTGCATATG AGAGAATGTG CAACATTCTC AAAGGGAAAT TCCAAACAGC 720

AGCACAAAGA GCAATGATGG ATCAGGTACG GGAAAGCAGA AATCCTGGGA ATGCTGAGAT 780CGAAGATCTC ATATTTCTGG CACGGTCTGC ACTCATCCTG AGAGGATCAG TGGCCCACAA 840GTCCTGCTTG CCTGCTTGTG TGTACGGGCT TGCCGTGGCC AGTGGATATG ACTTTGAGAG 900AGAAGGGTAC TCTCTGGTCG GGATTGATCC TTTCCGTCTG CTGCAAAACA GCCAGGTCTT 960TAGTCTAATT AGACCAAATG AGAATCCAGC ACATAAAAGT CAATTGGTGT GGATGGCATG 1020CCATTCTGCA GCATTTGAAG ATCTGAGAGT CTCAAGCTTC ATCAGAGGGA CAAGAGTGGC 1080CCCAAGGGGA CAACTATCTA CTAGAGGAGT ACAAATTGCT TCAAATGAGA ACATGGAAAC 1140AATGGACTCC AGCACTCTTG AACTGAGAAG CAGATATTGG GCTATAAGGA CCAGGAGTGG 1200AGGAAACACC AACCAGCAGA GAGCATCTGC AGGACAAATC AGTGTGCAGC CTACTTTCTC 1260GGTACAGAGA AATTCTTCCCT TCGAAAGAGC GACCATTATG GCGGCATTCA CAGGGAATAC 1320AGAGGGCAGA ACATCTGACA TGAGGACTGA AATCATAAGG ATGATGGAAA GCTCCAGACC 1380AGAAGGTCGTG TCTTTGACCAGGAGT G CAACGAA 1440CCCGATCGTG CCTTCCTTTG ACATGAGTAA TGAAGGATCT TATTTCTTCG GAGACAATGC 1500AGAGGAGTAT GACAATTGAA G 1521SEQ ID NO: 1454 PA: Consensus sequence derived from H5N1gi|47156500|AY585473|Influenza A virus (A/duck/Guangxi/35/2001(H5N1)) polymerase (PA) mRNA, complete coding sequence ATGGAAGACT TTGTGCGACA ATGCTTCAAT CCAATGATTG TCGAGCTTGC GGAAAAGGCA 60ATGAAAGAAT ATGGGGAAGA TCCGAAAATC GAAACGAACA AATTTGCAGC AATATGCACA 120CACTTAGAAG TCTGTTTCAT GTATTCAGAT TTTCACTTTA TTGATGAACG GGGCGAATCA 180ATAATTGTAG AATCTGGCGA TCCGAATGCA TTATTGAAAC ACCGATTTGA AATAATTGAA 240GGAAGAGACC GAACAATGGC CTGGACAGTG GTGAATAGTA TCTGCAACAC CACAGGAGTT 300GAGAAACCTA AATTTCTCCC AGATTTGTAT GACTACAAAG AGAACCGATT CATTGAAATT 360GGAGTGACAC GGAGGGAAGT TCATATATAC TATCTAGAGA AAGCCAACAA GATAAAATCC 420GAGAAGACAC ACATTCACAT ATTCTCATTC ACTGGGGAGG AAATGGCCAC CAAAGCGGAC 480TACACCCTTG ATGAAGAGAG CAGGGCAAGA ATCAAAACCA GGCTGTTCAC CATAAGGCAG 540GAAATGGCCA GTAGGGGTCT ATGGGATTCC TTTCGTCAGT CCGAGAGAGG CGAAGAGACA 600ATTGAAGAAA GATTTGAAAT CACAGGAACC ATGCGCAGGC TTGCCGACCA AAGTCTCCA 660CCGAACTTCT CCAGCCTTGA AAACTTTAGA GCCTATGTGG ATGGATTCGA ACCGAACGGC 720TGCATTGAGG GCAAGCTTTC TCAAATGTCA AAAGAAGTGA ACGCCAGAAT TGAGCCATTT 780CTGAAGACAA CACCACGCCC TCTCAGATTA CCTGATGGGC CTCCCTGCTC TCAGCGGTCG 840AAGTTCTTGC TGATGGATGC CCTTAAATTA AGCATCGAAG ACCCGAGTCA TGAGGGGGAG 900GGGATACCGC TATATGATGC AATCAAATGC ATGAAAACAT TTTTCGGCTG GAAAGAGCCC 960AACATCGTAA AACCACATGA AAAAGGCATA AACCCCAATT ACCTCCTGGC TTGGAAGCAA 1020GTGCTGGCAG AACTCCAAGA TATTGAAAAT GAGGAGAAAA TCCCAAAAAC AAAGAACATG 1080AAGAAAACAA GCCAATTGAA GTGGGCACTC GGTGAGAACA TGGCACCAGA GAAAGTAGAC 1140TTTGAGGATT GCAAAGATGT TAGCGATCTA AGACAGTATG ACAGTGATGA ACCAGAGCCT 1200AGATCACTAG CAAGCTGGAT CCAGAGTGAA TTCAACAAGG CATGTGAATT GACAGATTCG 1260AGTTGGATTG AACTTGATGA AATAGGGGAA GACGTTGCTC CAATTGAGCA CATTGCAAGT 1320ATGAGAAGTA ACTATTTCAC T ACCG CA GGGCCACTGA ATACATAATG 1380AAGGGGGTGT ACATAAACAC AGCTCTGTTG AATGCATCCT GTGCAGCCAT GGATGACTTT 1440CAACTGATTC CAATGATAAG CAAATGCAGA ACCAAAGAAG GAAGACGGAA AACTAACCTG 1500TATGGATTCA TTATAAAAGG AAGATCCCAT TTGAGGAATG ATACCGATGT GGTAAACTTT 1560GTGAGTATGG AATTCTCTCT TACTGACCCG AGGCTGGAGC CACACAAGTG GGAAAAGTAC 1620TGTGTTCTCG AGATAGGAGA CATGCTCCTA CGGACTGCAA TAGGCCAAGT TTCAAGGCCC 1680ATGTTCCTGT ATGTGAGAAC CAATGGAACC TCCAAGATCA AAATGAAATG GGGAATGGAG 1740ATGAGGCGAT GCCTTCTTCA ATCCCTTCAA CAGATTGAGA GCATGATTGA GGCCGAGTCT 1800

TCTGTCAAAG AGAAAGACAT GACCAAAGAA TTCTTTGAAA ACAAATCAGA AACATGGCCA 1860ATTGGAGAGT CACCCAAAGG AGTGGAGGAA GGCTCCATCG GGAAGGTGTG CAGAACCTTA 1920CTGGCGAAAT CTGTGTTCAA CAGTCTATAT GCATCTCCAC AACTCGAGGG GTTTTCAGCT 1980GAATCAAGAA AATTGCTTCT CATTGTTCAG GCACTTAGGG ACAACCTGGA ACCTGGGACC 2040TTCGATCTTG GAGGGCTATA TGAAGCAATT GAGGAGTGCC TGATTAATGA TCCCTGGGTT 2100TTGCTTAATG CGTCTTGGTT CAACTCCTTC CTCACACATG CACTGAAATA GTT 2153SEQ ID NO：1455

PB1: Derived from the viral consensus sequence of the PB1 gene of H5N1gi|58531084|AB166860|Influenza A virus (A/Chicken/Yamkou/7/2004(H5N1)) polymerase basic protein 1, complete coding sequence ATGGATGTCA ATCCGACTTT ACTTTTCTTG AAAGTACCAG TGCAAAATGC TATAAGTACC 60ACATTCCCTT ATACTGGAGA CCCTCCATAC AGCCATGGAA CAGGGACAGG ATACACCATG 120GACACAGTCA ACAGAACACA CCAATATTCA GAAAAGGGGA AGTGGACAAC AAACACAGAG 180ACTGGAGCAC CCCAACTCAA CCCGATTGAT GGACCACTAC CTGAGGATAA TGAGCCCTGT 240GGGTATGCAC AAACAGATTG TGTATTGGAA GCAATGGCTT TCCTTGAAGA ATCCCACCCA 300GGGATCTTTG AAAACTCGTG TCTTGAAACG ATGGAAATTG TTCAACAAAC AAGAGTGGAT 360AAACTGACCC AAGGTCGCCA GACCTATGAC TGGACATTGA ATAGAAACCA ACCGGCTGCA 420ACTGCTTTGG CCAACACTAT AGAAATCTTC AGATCGAACG GTCTAACAGC CAATGAATCG 480GGACGGCTAA TAGATTTCCT CAAGGATGTG ATGGAATCAA TGGATAAGGA AGAAATGGAG 540ATAACAACAC ATTTCCAGAG AAAGAGAAGA GTGAGGGACA ACATGACCAA GAAAATGGTC 600ACACAAAGAA CAATAGGGAA GAAAAAACAA AGGCTGAACA AAAAGGCTA CCTGATAA GA 660GCACTGACAC TGAACACAAT GACAAAAGAT GCAGAAAGAG GCAAATTGAA GAGGCGAGCA 720ATTGCAACAC CCGGAATGCA AATCAGAGGA TTCGTGTACT TTGTTGAAAC ACTAGCGAGG 780AGTATCTGTG AGAAACTTGA GCAATCTGGA CTCCCAGTCG GAGGGAATGA GAAGAAGGCT 840AAATTGGCAA ACGTCGTGAG GAAGATGATG ACTAACTCAC AAGATACTGA ACTCTCCTTT 900ACAATTACTG GAGACAATAC CAAATGGAAT GAGAATCAGA ATCCTAGGAT GTTTCTGGCA 960ATGATAACGT ACATCACAAG GAACCAGCCA GAATGGTTTC GGAATGTCTT AAGCATTGCC 1020CCTATAATGT TCTCAAACAA AATGGCGAGA TTAGGAAAAG GATACATGTT CGAAAGTAAG 1080AGCATGAAGT TACGAACACA AATACCAGCA GAAATGCTTG CAAACATTGA TCTCAAATAC 1140TTCAATGAAT TAACGAAAAA GAAAATTGAG AAAATAAGAC CTCTATTAAT AGATGGTACA 1200GCCTCATTGA GCCCTGGAAT GATGATGGGC ATGTTCAACA TGCTGAGTAC AGTCCTAGGA 1260GTCTCAATCC TGAATCTTGG ACAGAAAAGG TACACCAAAA CCACATATTG GTGCTGGACGGA 1320CTCCAATCCT CCTTC ATGCAC CGAATCATGA GGGAATACAA 1380GCAGGAGTGG ATAGGTTTTA TAGGACTTGT AAACTAGTTG GAATCAATAT GAGCAAGAAG 1440AAGTCTTACA TAAATCGGAC AGGGACATTT GAATTCACGA GCTTTTTCTA CCGCTATGGA 1500TTTGTAGCCA ATTTCAGTAT GGAGCTGCCC AGTTTTGGAG TGTCTGGAAT TAATGAATCG 1560GCCGACATGA GCATTGGTGT TACAGTGATA AAGAACAATA TGATAAACAA CGACCTTGGG 1620CCAGCAACAG CTCAGATGGC TCTTCAGCTA TTCATCAAGG ACTACAGATA CACATACCGA 1680TGCCACAGAG GGGATACGCA AATCCAAACG AGGAGATCAT TCGAGCTGAA GAAGCTGTGG 1740GAGCAAACCC GTTCAAAGGC AGGACTGTTG GTTTCAGATG GAGGACCAAA TCTATACAAT 1800ATCCGAAATC TCCATATTCC TGAGGTCTGC TTAAAATGGG AATTGATGGA TGAAGATTAC 1860CAGGGCAGAC TGTGTAATCC TCTGAATCCG TTCGTCAGCC ATAAGGAAAT TGAATCTGTC 1920AACAATGCTG TAGTAATGCC AGCTCATGGC CCGGCCAAAA GCGTGGAATA TGATGCCGTT 1980GCAACTACAC ATTCATGGAT TCCTAAAAGG AATCGTTCCA TTCTCAGAC G TCTTG AGGATGAACA GATGTACCAG AAGTGCTGCA ATCTATTCGA GAAATTCTTC 2100CCCAGCAGTT CATATCGGAG GCCAGTTGGA ATTTCCAGCA TGGTGGAGGC CATGGTGTCT 2160AGGGCCCGAA TTGACGCACG AATTGATTTC GAGTCTGGAA GGATTAAGAA AGAAGAGTTT 2220GCTGAGATCA TGAAGATCTG TTCCACCATT GAAGAGCTCA GACGGCAAAA ATAG 2274SEQ ID NO：1456 PB2: derived from H5N1gi|19697859|AY059525|Influenza A virus (A/duck/Hong Kong/2986.1/2000(H5N1)) fragment 1 polymerase (PB2) gene viral consensus sequence, partial coding sequence. ATGGAGAGAA TAAAAGAATT AAGAGATCTA ATGTCGCAGT CTCGCACTCG CGAGATACTA 60ACAAAAACCA CTGTGGACCA TATGGCCATA ATCAAGAAAT ACACATCAGG AAGACAAGAG 120AAGAACCCCTG CTCTCAGAAT GAAATGGATG ATGGCAATGA AATATCCAAT CACAGCAGAC 180

AAGAGAATAA TAGAGATGAT TCCTGAAAGG AATGAACAAG GGCAGACGCT TTGGAGCAAG 240ACAAATGATG CTGGATCGGA CAGGGTGATG GTGTCTCCCC TAGCTGTAAC TTGGTGGAAT 300AGGAATGGGC CGACGACAAG TGCAGTCTAT TATCCAAAGG TTTACAAAAC ATACTTTGAG 360AAGGTTGAAA GGTTAAAACA TGGAACCTTC GGTCCCGTTC ATTTCCGAAA CCAAATTAAA 420ATACGCCGCC GAGTTGATAT AAATCCTGGC CATGCAGATC TCAATGCTAA AGAAGCACAA 480GATGTCATCA TGGAGGTCGT TTTCCCAAAT GAAGTGGGAG CTAGAATATT GACATCAGAG 540TCGCAATTGA CAATAACGAA AGAAAAGAAA GAAGAGCTCC AGGATTGTAA GATTGCTCCT 600TTAATGGTTG CATACATGTT GGAAAGGGAA CTGGTCCGCA AAACCAGATT CCTACCGGTA 660GCAGGCGGAA CAAGCAGTGT GTACATTGAG GTATTGCATT TGACTCAAGG GACCTGCTGG 720GAACAGATGT ACACTCCAGG CGGAGAAGTG AGAAATGACG ATGTTATTCCA GAGTATGATC 780ATCGCTGCCA GAAACATTGT TAGGAGAGCA ACGGTATCAG CGGATCCACT GGCATCACTG 840CTGGAGATGTGTCAGATCACGAGAATAG CCTTAGGCAA 900AATCCAACTG AGGAACAAGC TGTGGATATA TGCAAAGCAG CAATGGGTTT GAGGATCAGT 960TCATCCTTTA GCTTTGGAGG CTTCACTTTC AAAAGAACAA GTGGAACATC CGTCAAGAAG 1020GAAGAGGAAG TGCTTACAGG CAACCTCCAA ACATTGAAAA TAAGAGTACA TGAGGGGTAT 1080GAGGAATTCA CAATGGTTGG GCGGAGGGCA ACAGCTATCC TGAGGAAAGC AACTAGAAGG 1140CTGATTCAGT TGATAGTAAG TGGAAGAGAC GAACAATCAA TCGCTGAGGC AATCATTGTA 1200GCAATGGTGT TCTCACAGGA GGATTGCATG ATAAAGGCAG TCCGAGGCGA TCTGAATTTC 1260GTAAACAGAG CAAACCAAAG ATTAAACCCC ATGCATCAAC TCCTGAGACA TTTTCAAAAG 1320GATGCAAAAG TGCTATTTCA GAATTGGGGA ATTGAACCCA TTGATAATGT CATGGGGATG 1380ATCGGAATAT TACCTGACAT GACTCCCAGC ACAGAAATGT CACTGAGAGG AGTAAGAGTT 1440AGTAAAATGG GAGTGGATGA ATATTCCAGC ACTGAGAGAG TAGTTGTAAG TATTGACCGT 1500TTCTTAAGGG TTCGAGATCA GCGGGGGAAC GTACTCTTAT CTCCCGAAGA GGTCAGCGAA 1560ACACAGGGAA AGAGAAATT GGCAATAACA TATTCATCAT CAATGATGTG GGAAATCAAC 1620GGTCCTGAGT CAGTGCTTGT TAACACCTAT CAATGGATCA TCAGAAACTG GGAGACTGTG 1680AAGATTCAAT GGTCTCAAGA CCCCACGATG CTGTACAATA AGATGGAGTT TGAACCGTTC 1740CAATCCTTGG TACCTAAAGC TGCCAGAGGT CAATACAGTG GATTTGTGAG AACACTATTC 1800CAACAAATGC GTGACGTACT GGGGACATTT GATACTGTCC AGATAATAAA GCTGCTACCA 1860TTTGCAGCAG CCCCACCGGA GCAGAGCAGA ATGCAGTTTT CTTCTCTAAC TGTGAATGTG 1920AGAGGCTCAG GAATGAGAAT ACTTGTAAGG GGCAATTCCC CTGTGTTCAA CTACAATAAG 1980GCAACCAAAA GGCTTACCGT TCTTGGAAAG GACGCAGGTG CATTAACAGA GGATCCAGAT 2040GAGGGAACAG CCGGAGTGGA ATCTGCAGTA CTGAGGGGAT TCCTAATTCT AGGCAAGGAG 2100GACAAAAGAT ATGGACCAGC ATTGAGCATC AATGAACTGA GCAATCTTGC GAAAGGGGAG 2160AAAGCTAATG TGCTGATAGG GCAAGGAGAC GTGGTGTTGG TAATGAAAACG GAAACGGGAC 2220TCCAGCAGATAC TTACTGAGCCAG TAATC CATCAATTAG 2280 SEQ ID NO: 1457

Tables 1A, C, D and E list, for selected exemplary reagents of the invention, the identifiers of the duplexes, the sequences of the sense and antisense strands, the target genes of the reagents, and the results obtained from the above assays. Tables 1B and H list the duplex identifier, the duplex identifier of the corresponding unmodified sequence, the sense strand, and the reverse sequence for selected exemplary reagents containing a modified nucleic acid group for degradation stability. The sequence of the sense strand and the target gene of the reagent, in which all nucleotide-containing pyrimidine bases contain a 2'-O-methyl group on the sense strand, and all at 5'-ca-3' or 5'-ua -Pyrimidine bases containing nucleotides in the context of the 3' sequence contain a 2'-O-methyl group in the antisense strand, except where the antisense strand is at 5'-ca-3' or 5'-ua-3' Those reagents that do not contain nucleotides in the sequence context where all uridines in the 5'-ug-3' sequence context are 2'-O-methyl-modified nucleotides in the antisense strand (e.g., AL- DP-2295, AL-DP-2301 and AL-DP-2302). For some particularly preferred RNAi agents of the invention, Table 2 lists the concentration at which 50% of the maximal inhibition is determined based on the dose response in Cos-7 cells engineered to express the influenza gene of.

Table 6

Sequences Used in Influenza A Virus Matrix Protein (MP) Analysis

AY180470 Influenza A virus strain A/quail/Nanchang/12-340/2000 (H1N1) matrix protein (M) gene, partial coding sequence (partialcds).

AY633213 Influenza A virus (A/mallard/Alberta/211/98(H1N1)) matrix protein (M) gene, complete coding sequence (complete cds).

AY664487 Influenza A virus (A/Mallard/Alberta/119/98(H1N1)) non-functional matrix protein mRNA, partial sequence.

M55476 Influenza A virus matrix protein (M1) gene, complete coding sequence, and M2 protein (M2) gene, complete coding sequence.

M55479 Influenza A virus matrix protein (M1) gene, complete coding sequence, and M2 protein (M2) gene, complete coding sequence.

M55480 Influenza A virus matrix protein (M1) gene, complete coding sequence, and M2 protein (M2) gene, complete coding sequence.

M63528 Influenza A virus (A/Turkey/Minnesota/166/81(H1N1)) membrane protein M1 and membrane protein M2 genes, complete coding sequence.

U49119 Influenza A virus matrix protein M1 and M2(M) genes, complete coding sequence.

M and M2 genes of matrix protein of Z26859 influenza A virus

Z26860 M and M2 genes of influenza A virus matrix protein

AY422021 Influenza A virus (A/duck/Hokkaido/95/01(H2N2)) matrix protein 1(M) gene, partial coding sequence.

M12699 Avian Influenza A/Duck/NY/6750/78 RNA fragment 7 encoding M1 and M2, complete coding sequence.

AF213915 Influenza A virus (A/chicken/Italy/5945/95(H3N2)) fragment 7 matrix protein (M) gene, partial coding sequence.

AY180498 Influenza A virus strain A/chicken/Nanchang/3-120/2001 (H3N2) matrix protein (M) gene, partial coding sequence.

AY664458 Influenza A virus (A/ruddy turnstone/Delaware/142/99(H3N2)) non-functional matrix protein mRNA, partial sequence.

AY769614 Influenza A virus (A/Turkey/Ohio/313053/04(H3N2)) matrix protein gene, partial coding sequence.

AY779257 Influenza A virus (A/Turkey/North Carolina/12344/03(H3N2)) matrix protein 2 (M) gene, partial coding sequence; and matrix protein 1 (M) gene, complete coding sequence.

AY779258 Influenza A virus (A/Turkey/Minnesota/764-2/03(H3N2)) matrix protein 2 (M) gene, partial coding sequence; and matrix protein 1 (M) gene, complete coding sequence.

AY862623 Influenza A virus (A/Chicken/Korea/S6/03(H3N2)) matrix protein (M) gene, complete coding sequence.

AY862624 Influenza A virus (A/duck/Korea/S7/03(H3N2)) matrix protein (M) gene, complete coding sequence.

AY862625 Influenza A virus (A/duck/Korea/S8/03(H3N2)) matrix protein (M) gene, complete coding sequence.

AY862626 Influenza A virus (A/duck/Korea/S9/03(H3N2)) matrix protein (M) gene, complete coding sequence.

AY862627 Influenza A virus (A/duck/Korea/S10/03(H3N2)) matrix protein (M) gene, complete coding sequence.

AY862628 Influenza A virus (A/pigeon/Korea/S11/03(H3N2)) matrix protein (M) gene, complete coding sequence.

Z26858 Influenza A virus matrix protein M and M2 genes

AB166865 Influenza A virus (A/Chicken/Yangkou/7/2004(H5N1)) M1 and M2 genes of matrix protein and membrane ion channel, complete coding sequence.

AB188819 Influenza A virus (A/Chicken/Ojta/8/2004(H5N1)) membrane ion channel 2 and M2 and M1 genes of matrix protein 1, the complete coding sequence.

AF509043 Influenza A virus (A/Chicken/Hong Kong/FY150/01(H5N1)) M1 protein (M1) gene, complete coding sequence.

AF509044 Influenza A virus (A/pheasant/Hong Kong/FY155/01(H5N1)) M1 protein (M1) gene, complete coding sequence.

AF509045 Influenza A virus (A/silky chicken/Hong Kong/SF189/01(H5N1)) M1 protein (M1) gene, complete coding sequence.

AF509046 Influenza A virus (A/quail/Hong Kong/SF203/01(H5N1)) M1 protein (M1) gene, complete coding sequence.

AF509047 Influenza A virus (A/pigeon/Hong Kong/SF215/01(H5N1)) M1 protein (M1) gene, complete coding sequence.

AF509048 Influenza A virus (A/chicken/Hong Kong/SF219/01(H5N1)) M1 protein (M1) gene, complete coding sequence.

AF509049 Influenza A virus (A/chicken/Hong Kong/715.5/01(H5N1)) M1 protein (M1) gene, complete coding sequence.

AF509050 Influenza A virus (A/Chicken/Hong Kong/751.1/01(H5N1)) M1 protein (M1) gene, complete coding sequence.

AF509051 Influenza A virus (A/Chicken/Hong Kong/822.1/01(H5N1)) M1 protein (M1) gene, complete coding sequence.

AF509052 Influenza A virus (A/Chicken/Hong Kong/829.2/01(H5N1)) M1 protein (M1) gene, complete coding sequence.

AF509053 Influenza A virus (A/Chicken/Hong Kong/830.2/01(H5N1)) M1 protein (M1) gene, complete coding sequence.

AF509054 Influenza A virus (A/Chicken/Hong Kong/858.3/01(H5N1)) M1 protein (M1) gene, complete coding sequence.

AF509055 Influenza A virus (A/chicken/Hong Kong/866.3/01(H5N1)) M1 protein (M1) gene, complete coding sequence.

AF509056 Influenza A virus (A/Chicken/Hong Kong/867.1/01(H5N1)) M1 protein (M1) gene, complete coding sequence.

AF509057 Influenza A virus (A/chicken/Hong Kong/879.1/01(H5N1)) M1 protein (M1) gene, complete coding sequence.

AF509058 Influenza A virus (A/Chicken/Hong Kong/873.3/01(H5N1)) M1 protein (M1) gene, complete coding sequence.

AF509059 Influenza A virus (A/Chicken/Hong Kong/876.1/01(H5N1)) M1 protein (M1) gene, complete coding sequence.

AF509060 Influenza A virus (A/Chicken/Hong Kong/891.1/01(H5N1)) M1 protein (M1) gene, complete coding sequence.

AF509061 Influenza A virus (A/Chicken/Hong Kong/893.2/01(H5N1)) M1 protein (M1) gene, complete coding sequence.

AF509062 Influenza A virus (A/goose/Hong Kong/76.1/01(H5N1)) M1 protein (M1) gene, complete coding sequence.

AF509063 Influenza A virus (A/goose/Hong Kong/ww100/01(H5N1)) M1 protein (M1) gene, complete coding sequence.

AF509064 Influenza A virus (A/duck/Hong Kong/573.4/01(H5N1)) M1 protein (M1) gene, complete coding sequence.

AF509065 Influenza A virus (A/duck/Hong Kong/646.3/01(H5N1)) M1 protein (M1) gene, complete coding sequence.

AY059506 Influenza A virus (A/goose/Hong Kong/ww26/2000(H5N1)) fragment 7 matrix protein (M) gene, partial coding sequence.

AY059507 Influenza A virus (A/goose/Hong Kong/ww28/2000(H5N1)) fragment 7 matrix protein (M) gene, partial coding sequence.

AY059508 Influenza A virus (A/duck/Hong Kong/ww381/2000(H5N1)) fragment 7 matrix protein (M) gene, partial coding sequence.

AY059509 Influenza A virus (A/duck/Hong Kong/ww461/2000(H5N1)) fragment 7 matrix protein (M) gene, partial coding sequence.

AY059510 Influenza A virus (A/goose/Hong Kong/ww491/2000(H5N1)) fragment 7 matrix protein (M) gene, partial coding sequence.

AY059511 Influenza A virus (A/duck/Hong Kong/2986.1/2000(H5N1)) fragment 7 matrix protein (M) gene, partial coding sequence.

AY059512 Influenza A virus (A/goose/Hong Kong/3014.8/2000(H5N1)) fragment 7 matrix protein (M) gene, partial coding sequence.

AY075029 Influenza A virus (A/chicken/Hong Kong/317.5/2001(H5N1)) matrix protein 1 and matrix protein 2 (M) genes, complete coding sequence.

AY075035 Influenza A virus (A/duck/Hong Kong/380.5/2001(H5N1)) matrix protein 1 and matrix protein 2 (M) genes, complete coding sequence.

AY221530 Influenza A virus (A/Chicken/Hong Kong/NT873.3/01-MB(H5N1)) matrix protein (M) gene, complete coding sequence.

AY221531 Influenza A virus (A/chicken/Hong Kong/NT873.3/01(H5N1)) matrix protein (M) gene, complete coding sequence.

AY221532 Influenza A virus (A/chicken/Hong Kong/FY150/01-MB(H5N1)) matrix protein (M) gene, complete coding sequence.

AY221533 Influenza A virus (A/Chicken/Hong Kong/FY150/01(H5N1)) matrix protein (M) gene, complete coding sequence.

AY221534 Influenza A virus (A/pheasant/Hong Kong/FY155/01-MB(H5N1)) matrix protein (M) gene, complete coding sequence.

AY221535 Influenza A virus (A/pheasant/Hong Kong/FY155/01(H5N1)) matrix protein (M) gene, complete coding sequence.

AY221536 Influenza A virus (A/chicken/Hong Kong/YU822.2/01-MB(H5N1)) matrix protein (M) gene, complete coding sequence.

AY221537 Influenza A virus (A/chicken/Hong Kong/YU822.2/01(H5N1)) matrix protein (M) gene, complete coding sequence.

AY221538 Influenza A virus (A/chicken/Hong Kong/YU562/01(H5N1)) matrix protein (M) gene, complete coding sequence.

AY518361 Influenza A virus (A/duck/China/E319-2/03(H5N1)) membrane ion channel M2 and matrix protein M1(M) genes, complete coding sequence.

AY575895 Influenza A virus (A/Gs/Hong Kong/739.2/02(H5N1)) matrix protein (M) gene, complete coding sequence.

AY575896 Influenza A virus (A/Eg/Hong Kong/757.3/02(H5N1)) matrix protein (M) gene, partial coding sequence.

AY575897 Influenza A virus (A/G.H/Hong Kong/793.1/02(H5N1)) matrix protein (M) gene, partial coding sequence.

AY575898 Influenza A virus (A/Dk/Hong Kong/821/02(H5N1)) matrix protein (M) gene, partial coding sequence.

AY575899 Influenza A virus (A/Ck/Hong Kong/31.4/02(H5N1)) matrix protein (M) gene, complete coding sequence.

AY575900 Influenza A virus (A/Ck/Hong Kong/61.9/02(H5N1)) matrix protein (M) gene, complete coding sequence.

AY575901 Influenza A virus (A/Ck/Hong Kong/YU777/02(H5N1)) matrix protein (M) gene, complete coding sequence.

AY575902 Influenza A virus (A/Ck/Hong Kong/96.1/02(H5N1)) matrix protein (M) gene, complete coding sequence.

AY575903 Influenza A virus (A/Ck/Hong Kong/409.1/02(H5N1)) matrix protein (M) gene, complete coding sequence.

AY575904 Influenza A virus (A/Ph/Hong Kong/sv674.15/02(H5N1)) matrix protein (M) gene, complete coding sequence.

AY585378 Influenza A virus (A/duck/Fujian/01/2002(H5N1)) matrix protein mRNA, complete coding sequence.

AY585379 Influenza A virus (A/duck/Fujian/13/2002(H5N1)) matrix protein mRNA, complete coding sequence.

AY585380 Influenza A virus (A/duck/Fujian/17/2001(H5N1)) matrix protein mRNA, complete coding sequence.

AY585381 Influenza A virus (A/duck/Fujian/19/2000(H5N1)) matrix protein mRNA, complete coding sequence.

AY585382 Influenza A virus (A/duck/Guangdong/01/2001(H5N1)) matrix protein mRNA, complete coding sequence.

AY585383 Influenza A virus (A/duck/Guangdong/07/2000(H5N1)) matrix protein mRNA, complete coding sequence.

AY585384 Influenza A virus (A/duck/Guangdong/12/2000(H5N1)) matrix protein mRNA, complete coding sequence.

AY585385 Influenza A virus (A/duck/Guangdong/22/2002(H5N1)) matrix protein mRNA, complete coding sequence.

AY585386 Influenza A virus (A/duck/Guangdong/40/2000(H5N1)) matrix protein mRNA, complete coding sequence.

AY585387 Influenza A virus (A/duck/Guangxi/07/1999(H5N1)) matrix protein mRNA, complete coding sequence.

AY585388 Influenza A virus (A/duck/Guangxi/22/2001(H5N1)) matrix protein mRNA, partial coding sequence.

AY585389 Influenza A virus (A/duck/Guangxi/35/2001(H5N1)) matrix protein mRNA, complete coding sequence.

AY585390 Influenza A virus (A/duck/Guangxi/53/2002(H5N1)) matrix protein mRNA, complete coding sequence.

AY585391 Influenza A virus (A/duck/Shanghai/08/2001(H5N1)) matrix protein mRNA, complete coding sequence.

AY585392 Influenza A virus (A/duck/Shanghai/13/2001(H5N1)) matrix protein mRNA, complete coding sequence.

AY585393 Influenza A virus (A/duck/Shanghai/35/2002(H5N1)) matrix protein mRNA, complete coding sequence.

AY585394 Influenza A virus (A/duck/Shanghai/37/2002(H5N1)) matrix protein mRNA, complete coding sequence.

AY585395 Influenza A virus (A/duck/Shanghai/38/2001(H5N1)) matrix protein mRNA, complete coding sequence.

AY585396 Influenza A virus (A/duck/Zhejiang/11/2000(H5N1)) matrix protein mRNA, complete coding sequence.

AY585397 Influenza A virus (A/duck/Zhejiang/52/2000(H5N1)) matrix protein mRNA, complete coding sequence.

AY585398 Influenza A virus (A/duck/Guangxi/50/2001(H5N1)) matrix protein mRNA, complete coding sequence.

AY590578 Influenza A virus (A/chicken/Nakorn-Patom/Thailand/CU-K2/2004(H5N1)) matrix protein M2 and matrix protein M1(M) genes, partial and complete coding sequences.

AY609315 Influenza A virus (A/chicken/Guangdong/174/04(H5N1)) fragment 7, complete sequence.

AY651374 Influenza A virus (A/Ck/Indonesia/BL/2003(H5N1)) membrane ion channel 2 (M) gene, partial coding sequence; and matrix protein 1 (M) gene, complete coding sequence.

AY651375 Influenza A virus (A/Dk/Indonesia/MS/2004(H5N1)) membrane ion channel 2 and matrix protein 1(M) genes, complete coding sequence.

AY651376 Influenza A virus (A/Ck/Indonesia/PA/2003(H5N1)) membrane ion channel 2 and matrix protein 1(M) genes, complete coding sequence.

AY651377 Influenza A virus (A/Ck/Indonesia/2A/2003(H5N1)) membrane ion channel 2 and matrix protein 1(M) genes, complete coding sequence.

AY651378 Influenza A virus (A/Ck/Indonesia/4/2004 (H5N1)) membrane ion channel 2 and matrix protein 1 (M) genes, complete coding sequence.

AY651379 Influenza A virus (A/Ck/Indonesia/5/2004 (H5N1)) membrane ion channel 2 and matrix protein 1 (M) genes, complete coding sequence.

AY651380 Influenza A virus (A/Ck/Thailand/1/2004(H5N1)) membrane ion channel 2 and matrix protein 1(M) genes, complete coding sequence.

AY651381 Influenza A virus (A/Ck/Thailand/73/2004(H5N1)) membrane ion channel 2 (M) gene, partial coding sequence; and matrix protein 1 (M) gene, complete coding sequence.

AY651382 Influenza A virus (A/Ck/Thailand/9.1/2004(H5N1)) membrane ion channel 2 (M) gene, partial coding sequence; and matrix protein 1 (M) gene, complete coding sequence.

AY651383 Influenza A virus (A/Qa/Thailand/57/2004 (H5N1)) membrane ion channel 2 and matrix protein 1 (M) genes, complete coding sequence.

AY651384 Influenza A virus (A/bird/Thailand/3.1/2004(H5N1)) membrane ion channel 2 (M) gene, partial coding sequence; and matrix protein 1 (M) gene, complete coding sequence.

AY651385 Influenza A virus (A/Dk/Thailand/71.1/2004(H5N1)) membrane ion channel 2 (M) gene, partial coding sequence; and matrix protein 1 (M) gene, complete coding sequence.

AY651386 Influenza A virus (A/Gs/Thailand/79/2004(H5N1)) membrane ion channel 2 (M) gene, partial coding sequence; and matrix protein 1 (M) gene, complete coding sequence.

AY651391 Influenza A virus (A/Ck/Vietnam/33/2004(H5N1)) membrane ion channel 2 and matrix protein 1 (M) genes, complete coding sequence.

AY651392 Influenza A virus (A/Ck/Vietnam/35/2004(H5N1)) membrane ion channel 2 and matrix protein 1 (M) genes, complete coding sequence.

AY651393 Influenza A virus (A/Ck/Vietnam/36/2004(H5N1)) membrane ion channel 2(M) gene, partial coding sequence; and matrix protein 1(M) gene, complete coding sequence.

AY651394 Influenza A virus (A/Ck/Vietnam/37/2004(H5N1)) membrane ion channel 2 and matrix protein 1 (M) genes, complete coding sequence.

AY651395 Influenza A virus (A/Ck/Vietnam/38/2004(H5N1)) membrane ion channel 2 and matrix protein 1 (M) genes, complete coding sequence.

AY651396 Influenza A virus (A/Ck/Vietnam/39/2004(H5N1)) membrane ion channel 2(M) gene, partial coding sequence; and matrix protein 1(M) gene, complete coding sequence.

AY651397 Influenza A virus (A/Ck/Vietnam/C57/2004(H5N1)) membrane ion channel 2(M) gene, partial coding sequence; and matrix protein 1(M) gene, complete coding sequence.

AY651398 Influenza A virus (A/Dk/Vietnam/11/2004(H5N1)) membrane ion channel 2 and matrix protein 1 (M) genes, complete coding sequence.

AY651399 Influenza A virus (A/Gf/Hong Kong/38/2002 (H5N1)) membrane ion channel 2 and matrix protein 1 (M) genes, partial coding sequence.

AY651400 Influenza A virus (A/Ck/Hong Kong/31.2/2002 (H5N1)) membrane ion channel 2 and matrix protein 1 (M) genes, complete coding sequence.

AY651401 Influenza A virus (A/Ck/Hong Kong/37.4/2002 (H5N1)) membrane ion channel 2 and matrix protein 1 (M) genes, complete coding sequence.

AY651402 Influenza A virus (A/SCk/Hong Kong/YU100/2002(H5N1)) membrane ion channel 2 and matrix protein 1 (M) genes, complete coding sequence.

AY651403 Influenza A virus (A/Ck/Hong Kong/YU22/2002(H5N1)) membrane ion channel 2 and matrix protein 1 (M) genes, complete coding sequence.

AY651404 Influenza A virus (A/Ck/Hong Kong/3176.3/2002 (H5N1)) membrane ion channel 2 and matrix protein 1 (M) genes, partial coding sequence.

AY651405 Influenza A virus (A/Ck/Hong Kong/3169.1/2002 (H5N1)) matrix protein 1 and membrane ion channel 2 (M) genes, part of the coding sequence.

AY651406 Influenza A virus (A/Ck/Hong Kong/FY157/2003(H5N1)) membrane ion channel 2 and matrix protein 1 (M) genes, complete coding sequence.

AY651407 Influenza A virus (A/Ck/Hong Kong/YU324/2003(H5N1)) membrane ion channel 2 and matrix protein 1 (M) genes, complete coding sequence.

AY651408 Influenza A virus (A/Ck/Hong Kong/2133.1/2003 (H5N1)) membrane ion channel 2 and matrix protein 1 (M) genes, partial coding sequence.

AY651409 Influenza A virus (A/Ck/Hong Kong/NT93/2003(H5N1)) membrane ion channel 2 (M) gene, partial coding sequence; and matrix protein 1 (M) gene, complete coding sequence.

AY651410 Influenza A virus (A/Ck/Hong Kong/SSP141/2003(H5N1)) membrane ion channel 2(M) gene, partial coding sequence; and matrix protein 1(M) gene, complete coding sequence.

AY651411 Influenza A virus (A/Ck/Hong Kong/WF157/2003(H5N1)) membrane ion channel 2 and matrix protein 1 (M) genes, complete coding sequence.

AY651412 Influenza A virus (A/Peregrine/Hong Kong/D0028/2004(H5N1)) membrane ion channel 2 (M) gene, partial coding sequence; and matrix protein 1 (M) gene, complete coding sequence.

AY651413 Influenza A virus (A/Red-billed Gull/Hong Kong/12.1/2003(H5N1)) membrane ion channel 2 and matrix protein 1 (M) genes, complete coding sequence.

AY651414 Influenza A virus (A/heron/Hong Kong/861.1/2002 (H5N1)) membrane ion channel 2 and matrix protein 1 (M) genes, complete coding sequence.

AY651415 Influenza A virus (A/Dove/Hong Kong/862.7/2002(H5N1)) membrane ion channel 2 and matrix protein 1 (M) genes, complete coding sequence.

AY651416 Influenza A virus (A/tree sparrow/Hong Kong/864/2002 (H5N1)) matrix protein 1 and membrane ion channel 2 (M) genes, part of the coding sequence.

AY651417 Influenza A virus (A/teal/China/2978.1/2002 (H5N1)) membrane ion channel 2 and matrix protein 1 (M) genes, partial coding sequence.

AY651418 Influenza A virus (A/Dk/Honduras(HN)/5806/2003(H5N1)) membrane ion channel 2(M) gene, partial coding sequence; and matrix protein 1(M) gene, complete coding sequence.

AY651419 Influenza A virus (A/Dk/ST/4003/2003(H5N1)) membrane ion channel 2(M) gene, partial coding sequence; and matrix protein 1(M) gene, complete coding sequence.

AY651420 Influenza A virus (A/Ck/ST/4231/2003(H5N1)) membrane ion channel 2(M) gene, partial coding sequence; and matrix protein 1(M) gene, complete coding sequence.

AY651421 Influenza A virus (A/Dk/YN/6255/2003(H5N1)) membrane ion channel 2 (M) gene, partial coding sequence; and matrix protein 1 (M) gene, complete coding sequence.

AY651422 Influenza A virus (A/Dk/YN/6445/2003(H5N1)) membrane ion channel 2 (M) gene, partial coding sequence; and matrix protein 1 (M) gene, complete coding sequence.

AY651423 Influenza A virus (A/Ck/Y/374/2004(H5N1)) membrane ion channel 2 (M) gene, partial coding sequence; and matrix protein 1 (M) gene, complete coding sequence.

AY651424 Influenza A virus (A/Dk/HN/101/2004(H5N1)) membrane ion channel 2(M) gene, partial coding sequence; and matrix protein 1(M) gene, complete coding sequence.

AY651425 Influenza A virus (A/Dk/HN/303/2004(H5N1)) membrane ion channel 2(M) gene, partial coding sequence; and matrix protein 1(M) gene, complete coding sequence.

AY651426 Influenza A virus (A/Ph/ST/44/2004(H5N1)) membrane ion channel 2 (M) gene, partial coding sequence; and matrix protein 1 (M) gene, complete coding sequence.

AY651427 Influenza A virus (A/Ck/YN/115/2004(H5N1)) membrane ion channel 2(M) gene, partial coding sequence; and matrix protein 1(M) gene, complete coding sequence.

AY653194 Influenza A virus (A/Chicken/Jilin/9/2004(H5N1)) fragment 7, complete sequence.

AY676045 Influenza A virus strain (A/duck/Hong Kong/821/02(H5N1)) membrane protein (M) gene, complete coding sequence.

AY676046 Influenza A virus strain (A/egret/Hong Kong/757.2/03(H5N1)) membrane protein (M) gene, complete coding sequence.

AY676047 Influenza A virus strain (A/chicken/Korea/ES/03(H5N1)) membrane protein (M) gene, complete coding sequence.

AY676048 Influenza A virus strain (A/duck/Korea/ESD1/03(H5N1)) membrane protein (M) gene, complete coding sequence.

AY684709 Influenza A virus (A/chicken/Hubei/327/2004(H5N1)) matrix protein 2 (M2) and matrix protein 1 (M1) genes, complete coding sequence.

AY737292 Influenza A virus (A/chicken/Guangdong/191/04(H5N1)) fragment 7, complete sequence.

AY737298 Influenza A virus (A/chicken/Guangdong/178/04(H5N1)) fragment 7, complete sequence.

AY737306 Influenza A virus (A/duck/Guangdong/173/04(H5N1)) fragment 7, complete sequence.

AY770077 Influenza A virus (A/chicken/Hubei/489/2004(H5N1)) matrix protein 2 (M2) and matrix protein 1 (M1) genes, complete coding sequence.

AY770998 Influenza A virus (A/Chicken/Ayutthaya/Thailand/CU-23/04(H5N1)) matrix protein gene, complete coding sequence.

AY818145 Influenza A virus (A/chicken/Vietnam/C58/04(H5N1)) matrix protein M1 gene, complete coding sequence.

AY818146 Influenza A virus (A/quail/Vietnam/36/04(H5N1)) matrix protein M1 gene, complete coding sequence.

AY856865 Influenza A virus (A/duck/Shandong/093/2004(H5N1)) fragment, complete sequence.

DQ055851 Influenza A virus (A/Chicken/Yunnan/K001/2004(H5N1)) matrix protein M1 gene, complete coding sequence.

AB189048 Influenza A virus (A/chicken/Kyoto/3/2004(H5N1)) membrane ion channel M2, M1 gene; M2, matrix protein 1, complete coding sequence.

AB189056 Influenza A virus (A/crow/Kyoto/53/2004(H5N1)) membrane ion channel M2, M1; M2, matrix protein 1, complete coding sequence.

AB189064 Influenza A virus (A/crow/Osaka/102/2004(H5N1)) membrane ion channel M2, M1 gene; M2, matrix protein 1, complete coding sequence,.

AF046082 Influenza A virus (A/chicken/Hong Kong/220/97(H5N1)) matrix protein 2 (M2) and matrix protein 1 (M1) genes, complete coding sequence.

AF098560 Influenza A virus (A/chicken/Hong Kong/258/97(H5N1)) M1 matrix protein (M) and M2 matrix protein (M) genes, partial coding sequence.

AF098561 Influenza A virus (A/chicken/Hong Kong/y388/97(H5N1)) M1 matrix protein (M) and M2 matrix protein (M) genes, partial coding sequence.

AF098562 Influenza A virus (A/Chicken/Hong Kong/728/97(H5N1)) M1 matrix protein (M) and M2 matrix protein (M) genes, partial coding sequence.

AF098563 Influenza A virus (A/Chicken/Hong Kong/786/97(H5N1)) M1 matrix protein (M) and M2 matrix protein (M) genes, partial coding sequence.

AF098564 Influenza A virus (A/chicken/Hong Kong/915/97(H5N1)) M1 matrix protein (M) and M2 matrix protein (M) genes, partial coding sequence.

AF098566 Influenza A virus (A/duck/Hong Kong/p46/97(H5N1)) M1 matrix protein (M) and M2 matrix protein (M) genes, partial coding sequence.

AF098567 Influenza A virus (A/duck/Hong Kong/y283/97(H5N1)) M1 matrix protein (M) and M2 matrix protein (M) genes, partial coding sequence.

AF098568 Influenza A virus (A/goose/Hong Kong/w355/97(H5N1)) M1 matrix protein (M) and M2 matrix protein (M) genes, partial coding sequence.

AF144306 Influenza A virus (A/goose/Guangdong/1/96(H5N1)) matrix protein M1 and M2(M) genes, or splice products, complete coding sequence.

AF216711 Influenza A virus (A/Environment/Hong Kong/437-4/99(H5N1)) matrix protein 1 and matrix protein 2 genes, complete coding sequence.

AF216719 Influenza A virus (A/Environment/Hong Kong/437-6/99(H5N1)) matrix protein 1 and matrix protein gene, complete coding sequence.

AF216727 Influenza A virus (A/environment/Hong Kong/437-8/99(H5N1)) matrix protein 1 and matrix protein 2 genes, complete coding sequence.

AF216735 Influenza A virus (A/environment/Hong Kong/437-10/99(H5N1)) matrix protein 1 and matrix protein 2 genes, complete coding sequence.

AF359560 Influenza A virus (A/goose/Guangdong/3/97(H5N1)) matrix protein 1 and matrix protein 2 (M) genes, complete coding sequence.

AF398429 Influenza A virus (A/goose/Hong Kong/385.3/2000 (H5N1)) matrix protein 1 (M) gene, partial coding sequence.

AF398430 Influenza A virus (A/goose/Hong Kong/385.5/2000 (H5N1)) matrix protein 1 (M) gene, partial coding sequence.

AF468843 Influenza A virus (A/duck/Anyang/AVL-1/2001(H5N1)) matrix protein 1 and matrix protein 2 genes, complete coding sequence.

AF509040 Influenza A virus (A/chicken/Hong Kong/FY77/01(H5N1)) M1 protein (M1) gene, complete coding sequence.

AF509041 Influenza A virus (A/chicken/Hong Kong/YU562/01(H5N1)) M1 protein (M1) gene, complete coding sequence.

AF509042 Influenza A virus (A/chicken/Hong Kong/YU563/01(H5N1)) M1 protein (M1) gene, complete coding sequence.

AF073180 Influenza A virus (A/chick/NJ/15086-3/94 (H7N3NSA)) matrix protein 1 (M1) and matrix protein 2 (M2) genes, complete coding sequence.

AF073197 Influenza A virus (A/Turkey/Oregon/71(H7N3NSB)) matrix protein 1 (M1) and matrix protein 2 (M2) genes, complete coding sequence.

AY664433 Influenza A virus (A/Turnstone/New Jersey/65/85(H7N3)) non-functional matrix protein mRNA, partial sequence.

AY677732 Influenza A virus (A/Chicken/British Columbia/CN7-3/04(H7N3)) matrix protein 1 (M1) gene, complete coding sequence.

AF073198 Influenza A virus (A/Turkey/Colorado/13356/91 (H7N3NSA)) matrix protein 1 (M1) and matrix protein 2 (M2) genes, complete coding sequence.

AF073200 Influenza A virus (A/quail/Arkansas/16309-7/94 (H7N3NSA)) matrix protein 1 (M1) and matrix protein 2 (M2) genes, complete coding sequence.

AF073201 Influenza A virus (A/Turkey/Utah/24721-10/95 (H7N3NSA)) matrix protein 1 (M1) and matrix protein 2 (M2) genes, complete coding sequence.

AJ627492 Influenza A virus (A/Turkey/Italy/214845/2002(H7N3)) Membrane protein 1 gene and Membrane protein 2 gene, genomic RNA.

AJ627497 Influenza A virus (A/Turkey/Italy/220158/2002(H7N3)) membrane protein 1 gene and protein 2 gene, genomic RNA.

AY241600 Influenza A virus (A/chicken/New York/12273-11/99(H7N3)) matrix protein 1 and matrix protein 2 genes, complete coding sequence.

AY241602 Influenza A virus (A/chicken/NY/14714-9/99(H7N3)) matrix protein 1 and matrix protein 2 genes, complete coding sequence.

AY241615 Influenza A virus (A/duck/NJ/117228-7/01(H7N3)) matrix protein 1 and matrix protein 2 genes, complete coding sequence.

AY241616 Influenza A virus (A/duck/PA/143585/01(H7N3)) matrix protein 1 and matrix protein 2 genes, complete coding sequence.

AY300975 Influenza A virus (A/Blue-winged Teal/TX/2/01(H7N3) membrane protein (M) gene, complete coding sequence.

AY303652 Influenza A virus (A/chicken/Chile/176822/02(H7N3)) matrix protein 1 and matrix protein 2 genes, complete coding sequence.

AY303653 Influenza A virus (A/chicken/Chile/4322/02(H7N3)) matrix protein 1 and matrix protein 2 genes, complete coding sequence.

AY303654 Influenza A virus (A/Chicken/Chile/4957/02(H7N3)) matrix protein 1 gene, complete coding sequence; and matrix protein 2 gene, partial coding sequence.

AY303655 Influenza A virus (A/chicken/Chile/4968/02(H7N3)) matrix protein 1 and matrix protein 2 genes, complete coding sequence.

AY303656 Influenza A virus (A/chicken/Chile/4977/02(H7N3)) matrix protein 1 and matrix protein 2 genes, complete coding sequence.

AY303657 Influenza A virus (A/Turkey/Chile/4418/02(H7N3)) matrix protein 1 gene, complete coding sequence; and matrix protein 2 gene, partial coding sequence.

AY586427 Influenza A virus (A/Turkey/Italy/214845/02(H7N3)) matrix protein gene, partial coding sequence.

AY586428 Influenza A virus (A/Turkey/Italy/220158/2002(H7N3)) matrix protein gene, partial coding sequence.

AY586429 Influenza A virus (A/Duck/Italy/43/01(H7N3)) matrix protein gene, partial coding sequence.

AY586430 Influenza A virus (A/Duck/Italy/33/01(H7N3)) matrix protein gene, partial coding sequence.

AY611525 Influenza A virus (A/chick/British Columbia/04(H7N3)) matrix protein 2 (M) and matrix protein 1 (M) genes, complete coding sequence.

AY646079 Influenza A virus (A/chick/British Columbia/GSC_human_B/04(H7N3)) matrix protein 2 and matrix protein 1 (M) genes, complete coding sequence.

AY648288 Influenza A virus (A/GSC_chick_B/British Columbia/04(H7N3)) matrix protein 2 (M) and matrix protein 1 (M) genes, complete coding sequence.

AY650271 Influenza A virus (A/GSC_chick/British Columbia/04(H7N3)) matrix protein 2 (M) and matrix protein 1 (M) genes, complete coding sequence.

AJ619676 Influenza A virus (A/Chicken/Germany/R28/03(H7N7)) membrane protein 1 M1 gene, genomic RNA.

AY340086 Influenza A virus (A/Netherlands/124/03(H7N7)) matrix protein gene, partial coding sequence.

AY340087 Influenza A virus (A/Netherlands/126/03(H7N7)) matrix protein gene, partial coding sequence.

AY340088 Influenza A virus (A/Netherlands/127/03(H7N7)) matrix protein gene, partial coding sequence.

AY340089 Influenza A virus (A/Netherlands/219/03(H7N7)) matrix protein gene, complete coding sequence.

AY340090 Influenza A virus (A/Netherlands/33/03(H7N7)) matrix protein gene, complete coding sequence.

AY340091 Influenza A virus (A/chicken/Netherlands/1/03(H7N7)) matrix protein gene, complete coding sequence.

AY664468 Influenza A virus (A/Turnstone/Delaware/134/99(H7N7)) non-functional matrix protein mRNA, partial sequence.

L37795 Influenza A/Chick/Brescia/1902(H7N7) matrix protein (M1) gene and transmembrane protein (M2) gene, complete coding sequence.

L37796 Influenza A/FPV/Waterfish (H7N7) matrix protein (M1) gene and transmembrane protein (M2) gene, complete coding sequence.

L37797 Influenza A/FPV/Weybridge (H7N7) matrix protein (M1) gene and transmembrane protein (M2) gene, complete coding sequence.

M23917 Influenza A/chicken/FPV/Weybridge(H7N7) M1 matrix protein gene, complete coding sequence.

M23921 Influenza A/chicken/FPV/Weybridge (H7N7) M2 matrix protein gene, complete coding sequence.

M38299 Influenza A/FPV/Weybridge (H7N7) matrix (M) protein (fragment 7) gene, complete coding sequence.

M63523 Influenza A virus (A/chicken/Victoria/1/85(H7N7)) membrane protein M1 and membrane protein M2 genes, complete coding sequence.

M63526 Influenza A virus (strain A/FPV/Waterhead/27(H7N7)) membrane protein M1 and membrane protein M2 genes, complete coding sequence.

AB049165 Influenza A virus (A/parakeet/Chiba/1/97(H9N2)) membrane ion channel, matrix protein M1, M2 gene, complete coding sequence.

AB049166 Influenza A virus (A/parakeet/Narita Airport/92A/98(H9N2)) membrane ion channel, M1 and M2 genes of matrix protein, complete coding sequence.

AF222671 Influenza A virus (A/Silkie/Hong Kong/SF44/99(H9N2)) fragment 7M1(M1) gene, partial coding sequence.

AF508684 Influenza A virus (A/ostrich/South Africa/9508103/95(H9N2)) fragment 7 matrix protein M1(M) gene, complete coding sequence.

AF508685 Influenza A virus (A/chicken/Pakistan/4/99(H9N2)) fragment 7 matrix protein M1(M) gene, partial coding sequence.

AF508686 Influenza A virus (A/chicken/Pakistan/5/99(H9N2)) fragment 7 matrix protein M1(M) gene, partial coding sequence.

AF508687 Influenza A virus (A/chicken/Germany/R45/98(H9N2)) fragment 7 matrix protein M1(M) gene, complete coding sequence.

AF508688 Influenza A virus (A/duck/Germany/113/95(H9N2)) segment 7 matrix protein M1(M) gene, complete coding sequence.

AF508689 Influenza A virus (A/chicken/Iran/11T/99(H9N2)) fragment 7 matrix protein M1(M) gene, partial coding sequence.

AF508690 Influenza A virus (A/chicken/Saudi Arabia/532/99(H9N2)) fragment 7 matrix protein M1(M) gene, partial coding sequence.

AF508691 Influenza A virus (A/pheasant/Ireland/PV18/97(H9N2)) segment 7 matrix protein M1(M) gene, complete coding sequence.

AF508692 Influenza A virus (A/chicken/Korea/99029/99(H9N2)) segment 7 matrix protein M1(M) gene, complete coding sequence.

AF508693 Influenza A virus (A/chicken/Beijing/8/98(H9N2)) segment 7 matrix protein M1(M) gene, complete coding sequence.

AF508694 Influenza A virus (A/chicken/Guangdong/10/00(H9N2)) segment 7 matrix protein M1(M) gene, complete coding sequence.

AF508695 Influenza A virus (A/chicken/Guangdong/11/97(H9N2)) segment 7 matrix protein M1(M) gene, complete coding sequence.

AF508696 Influenza A virus (A/chicken/Heilongjiang/10/97(H9N2)) segment 7 matrix protein M1(M) gene, complete coding sequence.

AF508697 Influenza A virus (A/chicken/Henan/62/00(H9N2)) fragment 7 matrix protein M1(M) gene, complete coding sequence.

AF508698 Influenza A virus (A/chicken/Ningxia/5/99(H9N2)) segment 7 matrix protein M1(M) gene, complete coding sequence.

AF508699 Influenza A virus (A/Chicken/Sichuan/5/97(H9N2)) fragment 7 matrix protein M1(M) gene, partial coding sequence.

AF508700 Influenza A virus (A/Chicken/Shandong/6/96(H9N2)) segment 7 matrix protein M1(M) gene, complete coding sequence.

AF508701 Influenza A virus (A/chicken/Shijiazhuang/2/99(H9N2)) fragment 7 matrix protein M1(M) gene, partial coding sequence.

AF508702 Influenza A virus (A/chicken/Shenzhen/9/97(H9N2)) segment 7 matrix protein M1(M) gene, complete coding sequence.

AF508703 Influenza A virus (A/duck/Nanjing/1/97(H9N2)) segment 7 matrix protein M1(M) gene, complete coding sequence.

AF508704 Influenza A virus (A/quail/Shanghai/8/96(H9N2)) segment 7 matrix protein M1(M) gene, complete coding sequence.

AF523482 Influenza A virus (A/duck/Shantou/1043/00(H9N2)) matrix protein (M) gene, complete coding sequence.

AF523483 Influenza A virus (A/duck/Shantou/2134/00(H9N2)) matrix protein (M) gene, complete coding sequence.

AF523484 Influenza A virus (A/Wild duck/Shantou/4808/01(H9N2)) matrix protein (M) gene, complete coding sequence.

AF523485 Influenza A virus (A/duck/Shantou/1042/00(H9N2)) matrix protein (M) gene, complete coding sequence.

AF523486 Influenza A virus (A/duck/Shantou/2143/00(H9N2)) matrix protein (M) gene, complete coding sequence.

AF523487 Influenza A virus (A/duck/Shantou/2144/00(H9N2)) matrix protein (M) gene, complete coding sequence.

AF523488 Influenza A virus (A/duck/Shantou/1881/00(H9N2)) matrix protein (M) gene, complete coding sequence.

AF523489 Influenza A virus (A/duck/Shantou/1796/00(H9N2)) matrix protein (M) gene, complete coding sequence.

AF523490 Influenza A virus (A/duck/Shantou/2102/00(H9N2)) matrix protein (M) gene, complete coding sequence.

AF523491 Influenza A virus (A/duck/Shantou/830/00(H9N2)) matrix protein (M) gene, complete coding sequence.

AF523492 Influenza A virus (A/duck/Shantou/2088/01(H9N2)) matrix protein (M) gene, complete coding sequence.

AF523493 Influenza A virus (A/duck/Shantou/1605/01(H9N2)) matrix protein (M) gene, complete coding sequence.

AF523494 Influenza A virus (A/duck/Hong Kong/610/79(H9N2)) matrix protein (M) gene, complete coding sequence.

AF523495 Influenza A virus (A/duck/Hong Kong/552/79(H9N2)) matrix protein (M) gene, complete coding sequence.

AF523496 Influenza A virus (A/duck/Hong Kong/289/78(H9N2)) matrix protein (M) gene, complete coding sequence.

AF523497 Influenza A virus (A/duck/Hong Kong/86/76(H9N2)) matrix protein (M) gene, complete coding sequence.

AF523498 Influenza A virus (A/duck/Hong Kong/366/78(H9N2)) matrix protein (M) gene, partial coding sequence.

AF536719 Influenza A virus (A/chicken/Beijing/1/95(H9N2)) non-functional matrix protein gene, partial sequence.

AF536720 Influenza A virus (A/chicken/Beijing/2/97(H9N2)) non-functional matrix protein gene, partial sequence.

AF536721 Influenza A virus (A/chicken/Beijing/3/99(H9N2)) non-functional matrix protein gene, partial sequence.

AF536722 Influenza A virus (A/Chicken/Guangdong/97(H9N2)) non-functional matrix protein gene, partial sequence.

AF536723 Influenza A virus (A/chicken/Hebei/1/96(H9N2)) non-functional matrix protein gene, partial sequence.

AF536724 Influenza A virus (A/chicken/Hebei/2/98(H9N2)) non-functional matrix protein gene, partial sequence.

AF536725 Influenza A virus (A/chicken/Hebei/3/98(H9N2)) non-functional matrix protein gene, partial sequence.

AF536726 Influenza A virus (A/chicken/Henan/98(H9N2)) non-functional matrix protein gene e, partial sequence.

AF536727 Influenza A virus (A/chicken/Liaoning/99(H9N2)) non-functional matrix protein gene, partial sequence.

AF536728 Influenza A virus (A/Chicken/Shandong/98(H9N2)) non-functional matrix protein gene, partial sequence.

AJ291398 Influenza A virus (A/Chicken/Pakistan/2/99(H9N2)) M1 gene of matrix protein 1 (exon 1) and M2 gene of matrix protein 2 (exons 1 and 2), genomic RNA

AJ427865 Influenza A virus (A/quail/Hong Kong/FY298/00(H9N2)) part of the matrix protein m gene, genomic RNA

AY180461 Influenza A virus strain A/pigeon/Nanchang/2-0461/2000 (H9N2) matrix protein (M) gene, partial coding sequence.

AY180462 Influenza A virus strain A/duck/Nanchang/11-290/2000 (H9N2) matrix protein (M) gene, partial coding sequence.

AY180463 Influenza A virus strain A/duck/Nanchang/11-197/2000 (H9N2) matrix protein (M) gene, partial coding sequence.

AY180464 Influenza A virus strain A/duck/Nanchang/11-392/2000 (H9N2) matrix protein (M) gene, partial coding sequence.

AY180477 Influenza A virus strain A/chicken/Nanchang/4-361/2001 (H9N2) matrix protein (M) gene, partial coding sequence.

AY180485 Influenza A virus strain A/pigeon/Nanchang/11-145/2000 (H9N2) matrix protein (M) gene, partial coding sequence.

AY180486 Influenza A virus strain A/duck/Nanchang/1-0070/2000 (H9N2) matrix protein (M) gene, partial coding sequence.

AY180489 Influenza A virus strain A/duck/Nanchang/10-389/2000 (H9N2) matrix protein (M) gene, partial coding sequence.

AY180490 Influenza A virus strain A/chicken/Nanchang/1-0016/2000 (H9N2) matrix protein (M) gene, partial coding sequence.

AY180492 Influenza A virus strain A/pigeon/Nanchang/7-058/2000 (H9N2) matrix protein (M) gene, partial coding sequence.

AY180495 Influenza A virus strain A/quail/Nanchang/2-0460/2000 (H9N2) matrix protein (M) gene, partial coding sequence.

AY180502 Influenza A virus strain A/chicken/Nanchang/4-010/2000 (H9N2) matrix protein (M) gene, partial coding sequence.

AY180504 Influenza A virus strain A/quail/Nanchang/4-040/2000 (H9N2) matrix protein (M) gene, partial coding sequence.

AY180506 Influenza A virus strain A/chicken/Nanchang/4-301/2001 (H9N2) matrix protein (M) gene, partial coding sequence.

AY180516 Influenza A virus strain A/duck/Nanchang/7-092/2000 (H9N2) matrix protein (M) gene, partial coding sequence.

AY180519 Influenza A virus strain A/Wild duck/Nanchang/2-0480/2000 (H9N2) matrix protein (M) gene, partial coding sequence.

AY253755 Influenza A virus (A/chicken/Shanghai/F/98(H9N2)) matrix protein M1 and membrane ion channel M2 genes, complete coding sequence.

AY496852 Influenza A virus (A/chicken/Mudanjiang/0823/2000(H9N2)) matrix protein (M1) mRNA, complete coding sequence.

AY633165 Influenza A virus (A/Mallard/Alberta/17/91(H9N2)) matrix protein (M) gene, complete coding sequence.

AY633277 Influenza A virus (A/Mallard/Alberta/321/88(H9N2)) matrix protein (M) gene, complete coding sequence.

AY633293 Influenza A virus (A/Mallard/Alberta/11/91(H9N2)) matrix protein (M) gene, complete coding sequence.

AY664464 Influenza A virus (A/shorebird/Delaware/276/99(H9N2)) nonfunctional matrix protein mRNA, partial sequence.

AY664679 Influenza A virus (A/chicken/Hong Kong/CSW153/03(H9N2)) membrane protein M2(M) gene, partial coding sequence; and protein M1(M) gene, complete coding sequence.

AY664680 Influenza A virus (A/chicken/Hong Kong/AP45/03(H9N2)) membrane protein M2(M) gene, partial coding sequence; and membrane protein M1(M) gene, complete coding sequence.

AY664681 Influenza A virus (A/chicken/Hong Kong/BD90/03(H9N2)) membrane protein M2(M) gene, partial coding sequence; and membrane protein M1(M) gene, complete coding sequence.

AY664682 Influenza A virus (A/chicken/Hong Kong/CSW291/03(H9N2)) membrane protein M2(M) gene, partial coding sequence; and membrane protein M1(M) gene, complete coding sequence.

AY664683 Influenza A virus (A/chicken/Hong Kong/CSW304/03(H9N2)) membrane protein M2(M) and membrane protein M1(M) genes, partial coding sequence.

AY664684 Influenza A virus (A/chicken/Hong Kong/FY23/03(H9N2)) membrane protein M2(M) gene, partial coding sequence; and membrane protein M1(M) gene, complete coding sequence.

AY664685 Influenza A virus (A/guineafowl/Hong Kong/NT101/03(H9N2)) membrane protein M2(M) gene, partial coding sequence; and membrane protein M1(M) gene, complete coding sequence.

AY664686 Influenza A virus (A/chicken/Hong Kong/NT142/03(H9N2)) membrane protein M2(M) gene, partial coding sequence; and membrane protein M1(M) gene, complete coding sequence.

AY664687 Influenza A virus (A/chicken/Hong Kong/SF1/03(H9N2)) membrane protein M2(M) gene, partial coding sequence; and membrane protein M1(M) gene, complete coding sequence.

AY664688 Influenza A virus (A/chicken/Hong Kong/SSP101/03(H9N2)) membrane protein M2(M) gene, partial coding sequence; and membrane protein M1(M) gene, complete coding sequence.

AY664689 Influenza A virus (A/chicken/Hong Kong/TP38/03(H9N2)) membrane protein M2(M) gene, partial coding sequence; and membrane protein M1(M) gene, complete coding sequence.

AY664690 Influenza A virus (A/chicken/Hong Kong/WF126/03(H9N2)) membrane protein M2(M) gene, partial coding sequence; and membrane protein M1(M) gene, complete coding sequence.

AY664691 Influenza A virus (A/pigeon/Hong Kong/WF53/03(H9N2)) membrane protein M2(M) gene, partial coding sequence; and membrane protein M1(M) gene, complete coding sequence.

AY664692 Influenza A virus (A/pheasant/Hong Kong/WF54/03(H9N2)) membrane protein M2(M) gene, partial coding sequence; and membrane protein M1(M) gene, complete coding sequence.

AY664693 Influenza A virus (A/Grizzly Chicken/Hong Kong/NT184/03(H9N2)) membrane protein M2(M) gene, partial coding sequence; and membrane protein M1(M) gene, complete coding sequence.

AY664694 Influenza A virus (A/chicken/Hong Kong/WF120/03(H9N2)) membrane protein M2(M) and membrane protein M1(M) genes, partial coding sequence.

AY664695 Influenza A virus (A/chicken/Hong Kong/NT366/03(H9N2)) membrane protein M2(M) gene, partial coding sequence; and membrane protein M1(M) gene, complete coding sequence.

AY664696 Influenza A virus (A/chicken/Hong Kong/SSP418/03(H9N2)) membrane protein M2(M) gene, partial coding sequence; and membrane protein M1(M) gene, complete coding sequence.

AY664697 Influenza A virus (A/chicken/Hong Kong/YU427/03(H9N2)) membrane protein M2(M) gene, partial coding sequence; and membrane protein M1(M) gene, complete coding sequence.

AY800234 Influenza A virus (A/Chicken/Korea/S1/2003(H9N2)) matrix protein (M) gene, complete coding sequence.

AY862614 Influenza A virus (A/Silkie/Korea/S3/03(H9N2)) matrix protein (M) gene, complete coding sequence.

AY862615 Influenza A virus (A/Chicken/Korea/S4/03(H9N2)) matrix protein (M) gene, complete coding sequence.

AY862616 Influenza A virus (A/Chicken/Korea/S5/03(H9N2)) matrix protein (M) gene, complete coding sequence.

AY862617 Influenza A virus (A/Chicken/Korea/S12/03(H9N2)) matrix protein (M) gene, complete coding sequence.

AY862618 Influenza A virus (A/duck/Korea/S13/03(H9N2)) matrix protein (M) gene, complete coding sequence.

AY862619 Influenza A virus (A/dove/Korea/S14/03(H9N2)) matrix protein (M) gene, partial coding sequence.

AY862620 Influenza A virus (A/Chicken/Korea/S15/03(H9N2)) matrix protein (M) gene, complete coding sequence.

AY862621 Influenza A virus (A/Chicken/Korea/S16/03(H9N2)) matrix protein (M) gene, complete coding sequence.

AY862622 Influenza A virus (A/Chicken/Korea/S18/03(H9N2)) matrix protein (M) gene, complete coding sequence.

AF156458 Influenza A virus (A/chicken/Hong Kong/G9/97(H9N2)) fragment 7 matrix protein M1 (M1) and matrix protein M2 (M2) genes, complete coding sequence.

AF156459 Influenza A virus (A/chicken/Hong Kong/G23/97(H9N2)) fragment 7 matrix protein M1 (M1) and matrix protein M2 (M2) genes, complete coding sequence.

AF156460 Influenza A virus (A/pigeon/Hong Kong/Y233/97(H9N2)) segment 7 matrix protein M1 (M1) and matrix protein M2 (M2) genes, complete coding sequence.

AF156461 Influenza A virus (A/duck/Hong Kong/Y280/97(H9N2)) fragment 7 matrix protein M1 (M1) and matrix protein M2 (M2) genes, complete coding sequence.

AF156462 Influenza A virus (A/duck/Hong Kong/Y439/97(H9N2)) fragment 7 matrix protein M1 (M1) and matrix protein M2 (M2) genes, complete coding sequence.

AF156463 Influenza A virus (A/quail/Hong Kong/G1/97(H9N2)) segment 7 matrix protein M1 (M1) gene, complete coding sequence; and matrix protein M2 (M2) gene, partial coding sequence.

AF156464 Influenza A virus (A/chicken/Hong Kong/739/94(H9N2)) segment 7 matrix protein M1 (M1) gene, complete coding sequence; and matrix protein M2 (M2) gene, partial coding sequence.

AF156465 Influenza A virus (A/quail/Hong Kong/AF157/92(H9N2)) segment 7 matrix protein M1 (M1) gene, complete coding sequence; and matrix protein M2 (M2) gene, partial coding sequence.

AF156466 Influenza A virus (A/chicken/Beijing/1/94(H9N2)) segment 7 matrix protein M1 (M1) gene, complete coding sequence; and matrix protein M2 (M2) gene, partial coding sequence.

AF156467 Influenza A virus (A/Chicken/Korea/38349-p96323/96(H9N2)) fragment 7 matrix protein M1 (M1) gene, complete coding sequence; and matrix protein M2 (M2) gene, partial coding sequence.

AF156468 Influenza A virus (A/chicken/Korea/25232-96006/96(H9N2)) fragment 7 matrix protein M1 (M1) gene, complete coding sequence; and matrix protein M2 (M2) gene, partial coding sequence.

AF156469 Influenza A virus (A/shorebird/Delaware/9/96(H9N2)) segment 7 matrix protein M1 (M1) gene, complete coding sequence; and matrix protein M2 (M2) gene, partial coding sequence.

AF156470 Influenza A virus (A/quail/Arkansas/29209-1/93(H9N2)) fragment 7 matrix protein M1 (M1) gene, complete coding sequence; and matrix protein M2 (M2) gene, partial coding sequence.

AF156471 Influenza A virus (A/Turkey/California/189/66(H9N2)) segment 7 matrix protein M1 (M1) gene, complete coding sequence; and matrix protein M2 (M2) gene, partial coding sequence.

AF203788 Influenza A virus (A/Chicken/Korea/MS96/96(H9N2)) matrix protein 1 mRNA, complete coding sequence; and matrix protein 2 mRNA, partial coding sequence.

AF222662 Influenza A virus (A/quail/Hong Kong/A17/99(H9N2)) fragment 7M1(M1) gene, partial coding sequence.

AF222663 Influenza A virus (A/pigeon/Hong Kong/FY6/99(H9N2)) fragment 7M1(M1) gene, partial coding sequence.

AF222664 Influenza A virus (A/chicken/Hong Kong/NT16/99(H9N2)) fragment 7M1(M1) gene, partial coding sequence.

AF222665 Influenza A virus (A/quail/Hong Kong/SSP10/99(H9N2)) fragment 7M1(M1) gene, partial coding sequence.

AF222666 Influenza A virus (A/pheasant/Hong Kong/SSP11/99(H9N2)) fragment 7M1(M1) gene, partial coding sequence.

AF222667 Influenza A virus (A/chicken/Hong Kong/FY20/99(H9N2)) fragment 7M1(M1) gene, partial coding sequence.

AF222668 Influenza A virus (A/chicken/Hong Kong/KC12/99(H9N2)) fragment 7M1(M1) gene, partial coding sequence.

AF222669 Influenza A virus (A/quail/Hong Kong/NT28/99(H9N2)) fragment 7M1(M1) gene, partial coding sequence.

AF222670 Influenza A virus (A/chicken/Hong Kong/SF2/99(H9N2)) fragment 7M1(M1) gene, partial coding sequence.

Sequences Used in Influenza A Nucleocapsid Protein (NP) Analysis

AF156415 Influenza A virus (A/Turkey/California/189/66(H9N2)) fragment 5 nucleoprotein (NP) gene, partial coding sequence.

AF523423 Influenza A virus (A/duck/Hong Kong/86/76(H9N2)) nucleocapsid protein (NP) gene, complete coding sequence.

AF523424 Influenza A virus (A/duck/Hong Kong/366/78(H9N2)) nucleocapsid protein (NP) gene, partial coding sequence.

AF523421 Influenza A virus (A/duck/Hong Kong/289/78(H9N2)) nucleocapsid protein (NP) gene, complete coding sequence.

AF523422 Influenza A virus (A/duck/Hong Kong/552/79(H9N2)) nucleocapsid protein (NP) gene, partial coding sequence.

AY633279 Influenza A virus (A/Mallard/Alberta/321/88(H9N2)) nucleoprotein (NP) gene, complete coding sequence.

AY633295 Influenza A virus (A/Mallard/Alberta/11/91(H9N2)) nucleoprotein (NP) gene, partial coding sequence.

AY633167 Influenza A virus (A/Mallard/Alberta/17/91(H9N2)) nucleoprotein (NP) gene, complete coding sequence.

AF156410 Influenza A virus (A/quail/Hong Kong/AF157/92(H9N2)) fragment 5 nucleoprotein (NP) gene, complete coding sequence.

AF156414 Influenza A virus (A/quail/Arkansas/29209-1/93(H9N2)) fragment 5 nucleoprotein (NP) gene, partial coding sequence.

AF156408 Influenza A virus (A/chicken/Hong Kong/739/94(H9N2)) fragment 5 nucleoprotein (NP) gene, complete coding sequence.

AF156409 Influenza A virus (A/chicken/Beijing/1/94(H9N2)) fragment 5 nucleoprotein (NP) gene, complete coding sequence.

AF536699 Influenza A virus (A/chicken/Beijing/1/95(H9N2)) nucleoprotein (NP) gene, partial coding sequence.

AF508596 Influenza A virus (A/ostrich/South Africa/9508103/95(H9N2)) fragment 5 nucleoprotein (NP) gene, partial coding sequence.

AF508600 Influenza A virus (A/duck/Germany/113/95(H9N2)) fragment 5 nucleoprotein (NP) gene, partial coding sequence.

AB020778 Influenza A virus nucleoprotein gene, complete coding sequence.

AF508613 Influenza A virus (A/Chicken/Shandong/6/96(H9N2)) fragment 5 nucleoprotein (NP) gene, partial coding sequence.

AF508617 Influenza A virus (A/quail/Shanghai/8/96(H9N2)) fragment 5 nucleoprotein (NP) gene, partial coding sequence.

AF536703 Influenza A virus (A/chicken/Hebei/1/96(H9N2)) nucleoprotein (NP) gene, partial coding sequence.

AF156411 Influenza A virus (A/chicken/Korea/38349-96323/96(H9N2)) fragment 5 nucleoprotein (NP) gene, complete coding sequence.

AF156412 Influenza A virus (A/Chicken/Korea/25232-96006/96(H9N2)) fragment 5 nucleoprotein (NP) gene, partial coding sequence.

M63779 Influenza A/FPV/Dobson/Netherlands/27(H7N7) nucleoprotein mRNA, complete coding sequence.

M63784 j Influenza A/Teal/Iceland/29/80 (H7N7) nucleoprotein mRNA, complete coding sequence.

AJ620352 Influenza A virus (A/chicken/Germany/R28/03(H7N7)) nucleoprotein NP gene, genome RNA.

AY342425 Influenza A virus (A/Netherlands/219/03(H7N7)) nucleocapsid protein gene, complete coding sequence.

AY342426 Influenza A virus (A/Netherlands/033/03(H7N7)) nucleocapsid protein gene, complete coding sequence.

AY342427 Influenza A virus (A/chicken/Netherlands/1/03(H7N7)) nucleocapsid protein gene, complete coding sequence.

AF156413 Influenza A virus (A/Shorebird/Delaware/9/96(H9N2)) fragment 5 nucleoprotein (NP) gene, partial coding sequence.

AF203787 Influenza A virus (A/Chicken/Korea/MS96/96(H9N2)) nucleoprotein mRNA, complete coding sequence.

AF156402 Influenza A virus (A/chicken/Hong Kong/G9/97(H9N2)) fragment 5 nucleoprotein (NP) gene, complete coding sequence.

AF156403 Influenza A virus (A/chicken/Hong Kong/G23/97(H9N2)) fragment 5 nucleoprotein (NP) gene, complete coding sequence.

AF156404 Influenza A virus (A/pigeon/Hong Kong/Y233/97(H9N2)) fragment 5 nucleoprotein (NP) gene, partial coding sequence.

AF156405 Influenza A virus (A/duck/Hong Kong/Y280/97(H9N2)) fragment 5 nucleoprotein (NP) gene, partial coding sequence.

AF156406 Influenza A virus (A/duck/Hong Kong/Y439/97(H9N2)) fragment 5 nucleoprotein (NP) gene, complete coding sequence.

AF156407 Influenza A virus (A/quail/Hong Kong/G1/97(H9N2)) fragment 5 nucleoprotein (NP) gene, partial coding sequence.

AF508612 Influenza A virus (A/Chicken/Sichuan/5/97(H9N2)) fragment 5 nucleoprotein (NP) gene, partial coding sequence.

AF536702 Influenza A virus (A/chicken/Guangdong/97(H9N2)) nucleoprotein (NP) gene, partial coding sequence.

AF536700 Influenza A virus (A/chicken/Beijing/2/97(H9N2)) nucleoprotein (NP) gene, partial coding sequence.

AF508615 Influenza A virus (A/chicken/Shenzhen/9/97(H9N2)) fragment 5 nucleoprotein (NP) gene, partial coding sequence.

AF508616 Influenza A virus (A/duck/Nanjing/1/97(H9N2)) fragment 5 nucleoprotein (NP) gene, partial coding sequence.

AB049161 Influenza A virus (A/parakeet/Chiba/1/97(H9N2)) nucleoprotein NP gene, complete coding sequence.

AF508603 Influenza A virus (A/pheasant/Ireland/PV18/97(H9N2)) fragment 5 nucleoprotein (NP) gene, partial coding sequence.

AF508607 Influenza A virus (A/chicken/Guangdong/11/97(H9N2)) fragment 5 nucleoprotein (NP) gene, partial coding sequence.

AF508609 Influenza A virus (A/chicken/Heilongjiang/10/97(H9N2)) fragment 5 nucleoprotein (NP) gene, partial coding sequence.

AF508608 Influenza A virus (A/chicken/Hebei/4/98(H9N2)) fragment 5 nucleoprotein (NP) gene, partial coding sequence.

AF508605 Influenza A virus (A/chicken/Beijing/8/98(H9N2)) fragment 5 nucleoprotein (NP) gene, partial coding sequence.

AF508599 Influenza A virus (A/chicken/Germany/R45/98(H9N2)) fragment 5 nucleoprotein (NP) gene, partial coding sequence.

AF536708 Influenza A virus (A/chicken/Shandong/98(H9N2)) nucleoprotein (NP) gene, partial coding sequence.

AY253753 Influenza A virus (A/chicken/Shanghai/F/98(H9N2)) nucleoprotein (NP) gene, complete coding sequence.

AF536704 Influenza A virus (A/chicken/Hebei/2/98(H9N2)) nucleoprotein (NP) gene, partial coding sequence.

AF536705 Influenza A virus (A/chicken/Hebei/3/98(H9N2)) nucleoprotein (NP) gene, partial coding sequence.

AF536706 Influenza A virus (A/chicken/Henan/98(H9N2)) nucleoprotein (NP) gene, partial coding sequence.

AB049162 Influenza A virus (A/parakeet/Narita/92A/98(H9N2)) nucleoprotein NP gene, complete coding sequence.

AF186270 Influenza A virus (A/quail/Hong Kong/NT28/99(H9N2)) fragment 5 nucleoprotein (NP) gene, partial coding sequence.

AF186271 Influenza A virus (A/Silkie chick/Hong Kong/SF43/99(H9N2)) fragment 5 nucleoprotein (NP) gene, partial coding sequence.

AF186272 Influenza A virus (A/chicken/Hong Kong/SF2/99(H9N2)) fragment 5 nucleoprotein (NP) gene, partial coding sequence.

AF222614 Influenza A virus (A/quail/Hong Kong/A17/99(H9N2)) fragment 5 nucleoprotein (NP) gene, partial coding sequence.

AF222615 Influenza A virus (A/pigeon/Hong Kong/FY6/99(H9N2)) fragment 5 nucleoprotein (NP) gene, partial coding sequence.

AF222616 Influenza A virus (A/chicken/Hong Kong/NT16/99(H9N2)) fragment 5 nucleoprotein (NP) gene, partial coding sequence.

AF222617 Influenza A virus (A/quail/Hong Kong/SSP10/99(H9N2)) fragment 5 nucleoprotein (NP) gene, partial coding sequence.

AF222618 Influenza A virus (A/pheasant/Hong Kong/SSP11/99(H9N2)) fragment 5 nucleoprotein (NP) gene, partial coding sequence.

AF536707 Influenza A virus (A/chicken/Liaoning/99(H9N2)) nucleoprotein (NP) gene, partial coding sequence.

AJ291394 Influenza A virus (A/chicken/Pakistan/2/99(H9N2)) nucleoprotein NP gene, genomic RNA.

AF536701 Influenza A virus (A/chicken/Beijing/3/99(H9N2)) nucleoprotein (NP) gene, partial coding sequence.

AF508611 Influenza A virus (A/chicken/Ningxia/5/99(H9N2)) fragment 5 nucleoprotein (NP) gene, partial coding sequence.

AF508614 Influenza A virus (A/chicken/Shijiazhuang/2/99 (H9N2)) fragment 5 nucleoprotein (NP) gene, partial coding sequence.

AF508604 Influenza A virus (A/Chicken/Korea/99029/99(H9N2)) fragment 5 nucleoprotein (NP) gene, partial coding sequence.

AF222619 Influenza A virus (A/chicken/Hong Kong/FY20/99(H9N2)) fragment 5 nucleoprotein (NP) gene, partial coding sequence.

AF222620 Influenza A virus (A/chicken/Hong Kong/KC12/99(H9N2)) fragment 5 nucleoprotein (NP) gene, partial coding sequence.

AF222621 Influenza A virus (A/Silkie/Hong Kong/SF44/99(H9N2)) fragment 5 nucleoprotein (NP) gene, partial coding sequence.

AF508601 Influenza A virus (A/chicken/Iran/11T/99(H9N2)) fragment 5 nucleoprotein (NP) gene, partial coding sequence.

AF508602 Influenza A virus (A/chick/Saudi Arabia/532/99(H9N2)) fragment 5 nucleoprotein (NP) gene, partial coding sequence.

AF508597 Influenza A virus (A/chicken/Pakistan/4/99(H9N2)) fragment 5 nucleoprotein (NP) gene, partial coding sequence.

AF508598 Influenza A virus (A/chicken/Pakistan/5/99(H9N2)) fragment 5 nucleoprotein (NP) gene, partial coding sequence.

AF508606 Influenza A virus (A/Chicken/Guangdong/10/00(H9N2)) fragment 5 nucleoprotein (NP) gene, partial coding sequence.

AF508610 Influenza A virus (A/chicken/Henan/62/00 (H9N2)) fragment 5 nucleoprotein (NP) gene, partial coding sequence.

AF523410 Influenza A virus (A/duck/Shantou/1043/00(H9N2)) nucleocapsid protein (NP) gene, partial coding sequence.

AF523411 Influenza A virus (A/duck/Shantou/2134/00(H9N2)) nucleocapsid protein (NP) gene, partial coding sequence.

AF523413 Influenza A virus (A/duck/Shantou/1042/00(H9N2)) nucleocapsid protein (NP) gene, partial coding sequence.

AF523415 Influenza A virus (A/duck/Shantou/2102/00(H9N2)) nucleocapsid protein (NP) gene, partial coding sequence.

AF523416 Influenza A virus (A/duck/Shantou/830/00(H9N2)) nucleocapsid protein (NP) gene, partial coding sequence.

AF523417 Influenza A virus (A/duck/Shantou/2144/00(H9N2)) nucleocapsid protein (NP) gene, partial coding sequence.

AF523419 Influenza A virus (A/duck/Shantou/2143/00(H9N2)) nucleocapsid protein (NP) gene, partial coding sequence.

AF523420 Influenza A virus (A/duck/Shantou/1881/00(H9N2)) nucleocapsid protein (NP) gene, partial coding sequence.

AJ427864 Influenza A virus (A/quail/Hong Kong/FY298/00(H9N2)) partial nucleoprotein np gene, genomic RNA

AY180525 Influenza A virus (A/pigeon/Nanchang/2-0461/2000(H9N2)) nucleoprotein (NP) gene, partial coding sequence.

AY180534 Influenza A virus strain A/duck/Nanchang/7-092/2000 (H9N2) nucleoprotein (NP) gene, partial coding sequence.

AY180537 Influenza A virus strain A/duck/Nanchang/11-392/2000 (H9N2) nucleoprotein (NP) gene, partial coding sequence.

AY180538 Influenza A virus (A/pigeon/Nanchang/11-145/2000(H9N2)) nucleoprotein (NP) gene, partial coding sequence.

AY180542 Influenza A virus strain A/duck/Nanchang/11-197/2000 (H9N2) nucleoprotein (NP) gene, partial coding sequence.

AY180544 Influenza A virus strain A/duck/Nanchang/11-290/2000 (H9N2) nucleoprotein (NP) gene, partial coding sequence.

AY180560 Influenza A virus (A/pigeon/Nanchang/7-058/2000(H9N2)) nucleoprotein (NP) gene, partial coding sequence.

AY180562 Influenza A virus strain A/chicken/Nanchang/4-010/2000 (H9N2) nucleoprotein (NP) gene, partial coding sequence.

AY180563 Influenza A virus strain A/quail/Nanchang/4-040/2000 (H9N2) nucleoprotein (NP) gene, partial coding sequence.

AY180564 Influenza A virus (A/wild duck/Nanchang/2-0480/2000(H9N2)) nucleoprotein (NP) gene, partial coding sequence.

AY180575 Influenza A virus (A/quail/Nanchang/2-0460/2000 (H9N2)) nucleoprotein (NP) gene, partial coding sequence.

AY496851 Influenza A virus (A/chicken/Mudanjiang/0823/2000(H9N2)) nucleoprotein (np) mRNA, complete coding sequence.

AY180581 Influenza A virus (A/chicken/Nanchang/1-0016/2000(H9N2)) nucleoprotein (NP) gene, partial coding sequence.

AY180583 Influenza A virus strain A/duck/Nanchang/10-389/2000 (H9N2) nucleoprotein (NP) gene, partial coding sequence.

AY180584 Influenza A virus strain A/duck/Nanchang/1-0070/2000 (H9N2) nucleoprotein (NP) gene, partial coding sequence.

AY768567 Influenza A virus (A/chicken/Korea/SNU0028/00(H9N2)) nucleocapsid protein (NP) gene, partial coding sequence.

AY768568 Influenza A virus (A/chicken/Korea/SNU0037/00(H9N2)) nucleocapsid protein (NP) gene, partial coding sequence.

AY768569 Influenza A virus (A/chicken/Korea/SNU0057/00(H9N2)) nucleocapsid protein (NP) gene, partial coding sequence.

AY768570 Influenza A virus (A/chicken/Korea/SNU0073/00(H9N2)) nucleocapsid protein (NP) gene, partial coding sequence.

AY768571 Influenza A virus (A/chicken/Korea/SNU0091/00(H9N2)) nucleocapsid protein (NP) gene, partial coding sequence.

AY768572 Influenza A virus (A/chicken/Korea/SNU0140/00(H9N2)) nucleocapsid protein (NP) gene, partial coding sequence.

AY768573 Influenza A virus (A/chicken/Korea/SNU0146/00(H9N2)) nucleocapsid protein (NP) gene, partial coding sequence.

AY768574 Influenza A virus (A/chicken/Korea/SNU1035C/00(H9N2)) nucleocapsid protein (NP) gene, partial coding sequence.

AY268949 Influenza A virus (A/chicken/Wangcheng/4/2001(H9N2)) nucleoprotein mRNA, complete coding sequence.

AY180578 Influenza A virus strain A/chicken/Nanchang/4-301/2001 (H9N2) nucleoprotein (NP) gene, partial coding sequence.

AY180551 Influenza A virus (A/chicken/Nanchang/4-361/2001(H9N2)) nucleoprotein (NP) gene, partial coding sequence.

AF523418 Influenza A virus (A/duck/Shantou/2088/01(H9N2)) nucleocapsid protein (NP) gene, partial coding sequence.

AF523414 Influenza A virus (A/duck/Shantou/1605/01(H9N2)) nucleocapsid protein (NP) gene, partial coding sequence.

AF523412 Influenza A virus (A/Wild duck/Shantou/4808/01(H9N2)) nucleocapsid protein (NP) gene, partial coding sequence.

AY800236 Influenza A virus (A/chicken/Korea/S1/2003(H9N2)) nucleoprotein (NP) gene, partial coding sequence.

AY862646 Influenza A virus (A/silky chick/Korea/S3/03(H9N2)) nucleocapsid protein (NP) gene, partial coding sequence.

AY862647 Influenza A virus (A/chicken/Korea/S4/03(H9N2)) nucleocapsid protein (NP) gene, partial coding sequence.

AY862648 Influenza A virus (A/chicken/Korea/S5/03(H9N2)) nucleocapsid protein (NP) gene, partial coding sequence.

AY862649 Influenza A virus (A/chicken/Korea/S12/03(H9N2)) nucleocapsid protein (NP) gene, partial coding sequence.

AY862650 Influenza A virus (A/duck/Korea/S13/03(H9N2)) nucleocapsid protein (NP) gene, partial coding sequence.

AY862651 Influenza A virus (A/pigeon/Korea/S14/03(H9N2)) nucleocapsid protein (NP) gene, partial coding sequence.

AY862652 Influenza A virus (A/chicken/Korea/S15/03(H9N2)) nucleocapsid protein (NP) gene, partial coding sequence.

AY862653 Influenza A virus (A/chicken/Korea/S16/03(H9N2)) nucleocapsid protein (NP) gene, partial coding sequence.

AY862654 Influenza A virus (A/chicken/Korea/S18/03(H9N2)) nucleocapsid protein (NP) gene, partial coding sequence.

AY664717 Influenza A virus (A/chicken/Hong Kong/CSW153/03(H9N2)) nucleoprotein (NP) gene, complete coding sequence.

AY664718 Influenza A virus (A/chicken/Hong Kong/AP45/03(H9N2)) nucleoprotein (NP) gene, complete coding sequence.

AY664719 Influenza A virus (A/chicken/Hong Kong/BD90/03(H9N2)) nucleoprotein (NP) gene, complete coding sequence.

AY664720 Influenza A virus (A/chicken/Hong Kong/CSW291/03(H9N2)) nucleoprotein (NP) gene, complete coding sequence.

AY664721 Influenza A virus (A/chicken/Hong Kong/CSW304/03(H9N2)) nucleoprotein (NP) gene, complete coding sequence.

AY664722 Influenza A virus (A/chicken/Hong Kong/FY23/03(H9N2)) nucleoprotein (NP) gene, complete coding sequence.

AY664723 Influenza A virus (A/Grizzly Chicken/Hong Kong/NT101/03(H9N2)) nucleoprotein (NP) gene, complete coding sequence.

AY664724 Influenza A virus (A/chicken/Hong Kong/NT142/03(H9N2)) nucleoprotein (NP) gene, complete coding sequence.

AY664725 Influenza A virus (A/chicken/Hong Kong/SF1/03(H9N2)) nucleoprotein (NP) gene, complete coding sequence.

AY664726 Influenza A virus (A/chicken/Hong Kong/SSP101/03(H9N2)) nucleoprotein (NP) gene, complete coding sequence.

AY664727 Influenza A virus (A/chicken/Hong Kong/TP38/03(H9N2)) nucleoprotein (NP) gene, complete coding sequence.

AY664728 Influenza A virus (A/chicken/Hong Kong/WF126/03(H9N2)) nucleoprotein (NP) gene, complete coding sequence.

AY664729 Influenza A virus (A/pigeon/Hong Kong/WF53/03(H9N2)) nucleoprotein (NP) gene, complete coding sequence.

AY664730 Influenza A virus (A/pheasant/Hong Kong/WF54/03(H9N2)) nucleoprotein (NP) gene, complete coding sequence.

AY664731 Influenza A virus (A/Grizzly Chicken/Hong Kong/NT184/03(H9N2)) nucleoprotein (NP) gene, complete coding sequence.

AY664732 Influenza A virus (A/chicken/Hong Kong/WF120/03(H9N2)) nucleoprotein (NP) gene, complete coding sequence.

AY664733 Influenza A virus (A/chicken/Hong Kong/NT366/03(H9N2)) nucleoprotein (NP) gene, partial coding sequence.

AY664734 Influenza A virus (A/chicken/Hong Kong/SSP418/03(H9N2)) nucleoprotein (NP) gene, partial coding sequence.

AY664735 Influenza A virus (A/chicken/Hong Kong/YU427/03(H9N2)) nucleoprotein (NP) gene, partial coding sequence.

AY788915 Influenza A virus (A/chicken/China/HSS2004(H9N2)) nucleoprotein (NP) gene, complete coding sequence.

AY586423 Influenza A virus (A/Duck/Italy/33/01(H7N3)) nucleoprotein gene, partial coding sequence.

AY586424 Influenza A virus (A/Duck/Italy/43/01(H7N3)) nucleoprotein gene, partial coding sequence.

AY586425 Influenza A virus (A/Turkey/Italy/220158/2002(H7N3)) nucleoprotein gene, partial coding sequence.

AY586426 Influenza A virus (A/Turkey/Italy/214845/02(H7N3)) nucleoprotein gene, partial coding sequence.

AJ627486 Influenza A virus (A/Turkey/Italy/214845/2002(H7N3)) nucleoprotein NP gene, genome RNA.

AJ627495 Influenza A virus (A/Turkey/Italy/220158/2002(H7N3)) nucleoprotein NP gene, genome RNA.

AY303658 Influenza A virus (A/Chicken/Chile/176822/02(H7N3)) nucleoprotein gene, complete coding sequence.

AY303659 Influenza A virus (A/Chicken/Chile/4957/02(H7N3)) nucleoprotein gene, complete coding sequence.

AY611527 Influenza A virus (A/chick/British Columbia/04(H7N3)) nucleoprotein (NP) gene, complete coding sequence.

AY646081 Influenza A virus (A/Chicken/British Columbia/GSC_Human_B/04(H7N3)) nucleoprotein (NP) gene, complete coding sequence.

AY648290 Influenza A virus (A/GSC_Chick_B/British Columbia/04(H7N3)) nucleoprotein (NP) gene, complete coding sequence.

AY650273 Influenza A virus (A/GSC_Chicken/British Columbia/04(H7N3)) nucleoprotein (NP) gene, complete coding sequence.

AF144303 Influenza A virus (A/goose/Guangdong/1/96(H5N1)) nucleocapsid protein (NP) gene, complete coding sequence.

AF046084 Influenza A virus (A/chicken/Hong Kong/220/97(H5N1)) nucleoprotein gene, complete coding sequence.

AF057293 Influenza A virus (A/Chicken/Hong Kong/258/97(H5N1)) nucleoprotein mRNA, complete coding sequence.

AF098617 Influenza A virus (A/chicken/Hong Kong/y388/97(H5N1)) nucleoprotein (NP) gene, complete coding sequence.

AF098618 Influenza A virus (A/chicken/Hong Kong/728/97(H5N1)) nucleoprotein (NP) gene, complete coding sequence.

AF098619 Influenza A virus (A/chicken/Hong Kong/786/97(H5N1)) nucleoprotein (NP) gene, complete coding sequence.

AF098620 Influenza A virus (A/chicken/Hong Kong/915/97(H5N1)) nucleoprotein (NP) gene, complete coding sequence.

AF098621 Influenza A virus (A/duck/Hong Kong/p46/97(H5N1)) nucleoprotein (NP) gene, complete coding sequence.

AF098622 Influenza A virus (A/duck/Hong Kong/y283/97(H5N1)) nucleoprotein (NP) gene, complete coding sequence.

AF098623 Influenza A virus (A/goose/Hong Kong/w355/97(H5N1)) nucleoprotein (NP) gene, complete coding sequence.

AF370122 Influenza A virus (A/goose/Guangdong/3/97(H5N1)) nucleoprotein gene, complete coding sequence.

AF216712 Influenza A virus (A/Environment/Hong Kong/437-4/99(H5N1)) nucleoprotein gene, complete coding sequence.

AF216720 Influenza A virus (A/Environment/Hong Kong/437-6/99(H5N1)) nucleoprotein gene, complete coding sequence.

AF216728 Influenza A virus (A/Environment/Hong Kong/437-8/99(H5N1)) nucleoprotein gene, complete coding sequence.

AF216736 Influenza A virus (A/Environment/Hong Kong/437-10/99(H5N1)) nucleoprotein gene, complete coding sequence.

AY585429 Influenza A virus (A/duck/Guangxi/07/1999(H5N1)) nucleoprotein (NP) mRNA, complete coding sequence.

AY585439 Influenza A virus (A/duck/Zhejiang/11/2000(H5N1)) nucleoprotein (NP) mRNA, complete coding sequence.

AY585440 Influenza A virus (A/duck/Zhejiang/52/2000(H5N1)) nucleoprotein (NP) mRNA, complete coding sequence.

AY585428 Influenza A virus (A/duck/Guangdong/40/2000(H5N1)) nucleoprotein (NP) mRNA, complete coding sequence.

AY585423 Influenza A virus (A/duck/Fujian/19/2000(H5N1)) nucleoprotein (NP) mRNA, complete coding sequence.

AY585425 Influenza A virus (A/duck/Guangdong/07/2000(H5N1)) nucleoprotein (NP) mRNA, complete coding sequence.

AY585426 Influenza A virus (A/duck/Guangdong/12/2000(H5N1)) nucleoprotein (NP) mRNA, complete coding sequence.

AY059492 Influenza A virus (A/goose/Hong Kong/ww26/2000(H5N1)) fragment 5 nucleocapsid protein (NP) gene, partial coding sequence.

AY059493 Influenza A virus (A/goose/Hong Kong/ww28/2000(H5N1)) fragment 5 nucleocapsid protein (NP) gene, partial coding sequence.

AY059494 Influenza A virus (A/duck/Hong Kong/ww381/2000(H5N1)) fragment 5 nucleocapsid protein (NP) gene, partial coding sequence.

AY059495 Influenza A virus (A/duck/Hong Kong/ww461/2000(H5N1)) fragment 5 nucleocapsid protein (NP) gene, partial coding sequence.

AY059496 Influenza A virus (A/goose/Hong Kong/ww491/2000(H5N1)) fragment 5 nucleocapsid protein (NP) gene, partial coding sequence.

AY059497 Influenza A virus (A/duck/Hong Kong/2986.1/2000(H5N1)) fragment 5 nucleocapsid protein (NP) gene, partial coding sequence.

AY059498 Influenza A virus (A/goose/Hong Kong/3014.8/2000(H5N1)) fragment 5 nucleocapsid protein (NP) gene, partial coding sequence.

AF398419 Influenza A virus (A/goose/Hong Kong/385.3/2000(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AF398420 Influenza A virus (A/goose/Hong Kong/385.5/2000(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AF468842 Influenza A virus (A/duck/Anyang/AVL-1/2001(H5N1)) nucleoprotein (NP) gene, complete coding sequence.

AF509117 Influenza A virus (A/chicken/Hong Kong/FY77/01(H5N1)) nucleocapsid protein (NP) gene, complete coding sequence.

AF509118 Influenza A virus (A/chicken/Hong Kong/YU562/01(H5N1)) nucleocapsid protein (NP) gene, complete coding sequence.

AF509119 Influenza A virus (A/chicken/Hong Kong/YU563/01(H5N1)) nucleocapsid protein (NP) gene, complete coding sequence.

AY585438 Influenza A virus (A/duck/Shanghai/38/2001(H5N1)) nucleoprotein (NP) mRNA, complete coding sequence.

AY221548 Influenza A virus (A/chicken/Hong Kong/NT873.3/01-MB(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY221549 Influenza A virus (A/chicken/Hong Kong/NT873.3/01(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY221550 Influenza A virus (A/chicken/Hong Kong/FY150/01-MB(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY221551 Influenza A virus (A/chicken/Hong Kong/FY150/01(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY221552 Influenza A virus (A/pheasant/Hong Kong/FY155/01-MB(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY221553 Influenza A virus (A/pheasant/Hong Kong/FY155/01(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY221554 Influenza A virus (A/chicken/Hong Kong/YU822.2/01-MB(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY221555 Influenza A virus (A/chicken/Hong Kong/YU822.2/01(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY221556 Influenza A virus (A/chicken/Hong Kong/YU562/01(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AF509120 Influenza A virus (A/chicken/Hong Kong/FY150/01(H5N1)) nucleocapsid protein (NP) gene, complete coding sequence.

AF509121 Influenza A virus (A/pheasant/Hong Kong/FY155/01(H5N1)) nucleocapsid protein (NP) gene, complete coding sequence.

AF509122 Influenza A virus (A/Silky/Hong Kong/SF189/01(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AF509123 Influenza A virus (A/quail/Hong Kong/SF203/01(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AF509124 Influenza A virus (A/pigeon/Hong Kong/SF215/01(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AF509125 Influenza A virus (A/chicken/Hong Kong/SF219/01(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AF509126 Influenza A virus (A/chicken/Hong Kong/715.5/01(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AF509127 Influenza A virus (A/chicken/Hong Kong/751.1/01(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AF509128 Influenza A virus (A/chicken/Hong Kong/822.1/01(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AF509129 Influenza A virus (A/chicken/Hong Kong/829.2/01(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AF509130 Influenza A virus (A/chicken/Hong Kong/830.2/01(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AF509131 Influenza A virus (A/chicken/Hong Kong/858.3/01(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AF509132 Influenza A virus (A/chicken/Hong Kong/866.3/01(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AF509133 Influenza A virus (A/chicken/Hong Kong/867.1/01(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AF509134 Influenza A virus (A/chicken/Hong Kong/879.1/01(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AF509135 Influenza A virus (A/chicken/Hong Kong/873.3/01(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AF509136 Influenza A virus (A/chicken/Hong Kong/876.1/01(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AF509137 Influenza A virus (A/chicken/Hong Kong/891.1/01(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AF509138 Influenza A virus (A/chicken/Hong Kong/893.2/01(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AF509139 Influenza A virus (A/goose/Hong Kong/76.1/01(H5N1)) nucleocapsid protein (NP) gene, complete coding sequence.

AF509140 Influenza A virus (A/goose/Hong Kong/ww100/01(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AF509141 Influenza A virus (A/duck/Hong Kong/573.4/01(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AF509142 Influenza A virus (A/duck/Hong Kong/646.3/01(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY585424 Influenza A virus (A/duck/Guangdong/01/2001(H5N1)) nucleoprotein (NP) mRNA, complete coding sequence.

AY585422 Influenza A virus (A/duck/Fujian/17/2001(H5N1)) nucleoprotein (NP) mRNA, complete coding sequence.

AY585430 Influenza A virus (A/duck/Guangxi/22/2001(H5N1)) nucleoprotein (NP) mRNA, complete coding sequence.

AY585431 Influenza A virus (A/duck/Guangxi/35/2001(H5N1)) nucleoprotein (NP) mRNA, complete coding sequence.

AY585432 Influenza A virus (A/duck/Guangxi/50/2001(H5N1)) nucleoprotein (NP) mRNA, complete coding sequence.

AY585434 Influenza A virus (A/duck/Shanghai/08/2001(H5N1)) nucleoprotein (NP) mRNA, complete coding sequence.

AY585435 Influenza A virus (A/duck/Shanghai/13/2001(H5N1)) nucleoprotein (NP) mRNA, complete coding sequence.

AY585436 Influenza A virus (A/duck/Shanghai/35/2002(H5N1)) nucleoprotein (NP) mRNA, complete coding sequence.

AY585437 Influenza A virus (A/duck/Shanghai/37/2002(H5N1)) nucleoprotein (NP) mRNA, complete coding sequence.

AY585433 Influenza A virus (A/duck/Guangxi/53/2002(H5N1)) nucleoprotein (NP) mRNA, complete coding sequence.

AY585427 Influenza A virus (A/duck/Guangdong/22/2002(H5N1)) nucleoprotein (NP) mRNA, complete coding sequence.

AY585420 Influenza A virus (A/duck/Fujian/01/2002(H5N1)) nucleoprotein (NP) mRNA, complete coding sequence.

AY585421 Influenza A virus (A/duck/Fujian/13/2002(H5N1)) nucleoprotein (NP) mRNA, complete coding sequence.

AY575907 Influenza A virus (A/Gs/Hong Kong/739.2/02(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY575908 Influenza A virus (A/Eg/Hong Kong/757.3/02(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY575909 Influenza A virus (A/G.H/Hong Kong/793.1/02(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY575910 Influenza A virus (A/Dk/Hong Kong/821/02(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY575911 Influenza A virus (A/Ck/Hong Kong/31.4/02(H5N1)) nucleocapsid protein (NP) gene, complete coding sequence.

AY575912 Influenza A virus (A/Ck/Hong Kong/61.9/02(H5N1)) nucleocapsid protein (NP) gene, complete coding sequence.

AY575913 Influenza A virus (A/Ck/Hong Kong/YU777/02(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY575914 Influenza A virus (A/Ck/Hong Kong/96.1/02(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY575915 Influenza A virus (A/Ck/Hong Kong/409.1/02(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY575916 Influenza A virus (A/Ph/Hong Kong/sv674.15/02(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

DQ023146 Influenza A virus (A/chicken/sd/1/02(H5N1)) nucleoprotein (NP) mRNA, complete coding sequence.

AY676037 Influenza A virus (A/duck/Hong Kong/821/02(H5N1)) nucleoprotein (NP) gene, complete coding sequence.

AY651510 Influenza A virus (A/Gf/Hong Kong/38/2002(H5N1)) nucleocapsid protein (NP) gene, complete coding sequence.

AY651511 Influenza A virus (A/Ck/Hong Kong/31.2/2002(H5N1)) nucleocapsid protein (NP) gene, complete coding sequence.

AY651512 Influenza A virus (A/Ck/Hong Kong/37.4/2002(H5N1)) nucleocapsid protein (NP) gene, complete coding sequence.

AY651513 Influenza A virus (A/SCk/Hong Kong/YU100/2002(H5N1)) nucleocapsid protein (NP) gene, complete coding sequence.

AY651514 Influenza A virus (A/Ck/Hong Kong/YU22/2002(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY651521 Influenza A virus (A/Ck/Hong Kong/3176.3/2002(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY651522 Influenza A virus (A/Ck/Hong Kong/3169.1/2002 (H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY651524 Influenza A virus (A/Dove/Hong Kong/862.7/2002(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY651525 Influenza A virus (A/Tree Finch/Hong Kong/864/2002(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY651526 Influenza A virus (A/heron/Hong Kong/861.1/2002 (H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY651527 Influenza A virus (A/teal/China/2978.1/2002(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY651523 Influenza A virus (A/Red-billed Gull/Hong Kong/12.1/2003(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY651487 Influenza A virus (A/Ck/Indonesia/PA/2003(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY651515 Influenza A virus (A/Ck/Hong Kong/2133.1/2003(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY651516 Influenza A virus (A/Ck/Hong Kong/NT93/2003(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY651517 Influenza A virus (A/Ck/Hong Kong/SSP141/2003(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY651518 Influenza A virus (A/Ck/Hong Kong/WF157/2003(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY651519 Influenza A virus (A/Ck/Hong Kong/FY157/2003(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY651520 Influenza A virus (A/Ck/Hong Kong/YU324/2003(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY651490 Influenza A virus (A/Ck/Indonesia/2A/2003(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY676038 Influenza A virus (A/egret/Hong Kong/757.2/03(H5N1)) nucleoprotein (NP) gene, complete coding sequence.

AY676039 Influenza A virus (A/chicken/Korea/ES/03(H5N1)) nucleoprotein (NP) gene, complete coding sequence.

AY676040 Influenza A virus (A/duck/Korea/ESD1/03(H5N1)) nucleoprotein (NP) gene, complete coding sequence.

AY651529 Influenza A virus (A/Dk/HN/5806/2003(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY651532 Influenza A virus (A/Ck/ST/4231/2003 (H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY651534 Influenza A virus (A/Dk/ST/4003/2003(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY651535 Influenza A virus (A/Dk/YN/6255/2003(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY651536 Influenza A virus (A/Dk/YN/6445/2003(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY651485 Influenza A virus (A/Ck/Indonesia/BL/2003(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY518364 Influenza A virus (A/duck/China/E319-2/03(H5N1)) nucleocapsid protein (NP) gene, complete coding sequence.

AY574189 Influenza A virus (A/chicken/Vietnam/HD1/2004(H5N1)) nucleoprotein gene, partial coding sequence.

AY574192 Influenza A virus (A/chicken/Vietnam/HD2/2004(H5N1)) nucleoprotein gene, partial coding sequence.

AJ867076 Influenza A virus (A/Hatay/2004/(H5N1)) nucleoprotein NP gene, genomic RNA

AY651486 Influenza A virus (A/Dk/Indonesia/MS/2004(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY590579 Influenza A virus (A/chicken/Nakhon Pathom/Thailand/CU-K2/2004(H5N1)) nucleocapsid protein (NP) gene, complete coding sequence.

AY609313 Influenza A virus (A/chicken/Guangdong/174/04(H5N1)) fragment 5, complete sequence.

AY576929 Influenza A virus (A/chicken/Vietnam/CM/2004(H5N1)) fragment 5 nucleoprotein gene, partial coding sequence.

AY576931 Influenza A virus (A/muscovy duck/Vietnam/MdGL/2004(H5N1)) fragment 5 nucleoprotein gene, partial coding sequence.

AB166863 Influenza A virus (A/Chicken/Yamaguchi/7/2004(H5N1)) nucleoprotein NP gene, complete coding sequence.

AB188817 Influenza A virus (A/chicken/Oita/8/2004(H5N1)) nucleoprotein NP gene, complete coding sequence.

AY651537 Influenza A virus (A/Ck/YN/374/2004(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY651538 Influenza A virus (A/Ck/YN/115/2004(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY653196 Influenza A virus (A/Chicken/Jilin/9/2004(H5N1)) fragment 5, complete sequence.

AY651533 Influenza A virus (A/Ph/ST/44/2004(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY651530 Influenza A virus (A/Dk/HN/303/2004(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY651531 Influenza A virus (A/Dk/HN/101/2004(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY684707 Influenza A virus (A/chicken/Hubei/327/2004(H5N1)) nucleoprotein (NP) gene, complete coding sequence.

AY737290 Influenza A virus (A/chicken/Guangdong/191/04(H5N1)) fragment 5, complete sequence.

AY737297 Influenza A virus (A/chicken/Guangdong/178/04(H5N1)) fragment 5, complete sequence.

AY737305 Influenza A virus (A/duck/Guangdong/173/04(H5N1)) fragment 5, complete sequence.

AY770081 Influenza A virus (A/chicken/Hubei/489/2004(H5N1)) nucleoprotein (NP) gene, complete coding sequence.

AY770996 Influenza A virus (A/chicken/Ayutthaya/Thailand/CU-23/04(H5N1)) nucleoprotein gene, partial coding sequence.

AY818139 Influenza A virus (A/chicken/Vietnam/C58/04(H5N1)) nucleoprotein NP gene, complete coding sequence.

AY818140 Influenza A virus (A/quail/Vietnam/36/04(H5N1)) nucleoprotein NP gene, complete coding sequence.

AY856864 Influenza A virus (A/duck/Shandong/093/2004(H5N1)) fragment 5, complete sequence.

AB189046 Influenza A virus (A/chicken/Kyoto/3/2004(H5N1)) nucleoprotein NP gene, complete coding sequence, .

AB189054 Influenza A virus (A/crow/Kyoto/53/2004(H5N1)) nucleoprotein NP gene, complete coding sequence, .

AB189062 Influenza A virus (A/crow/Osaka/102/2004(H5N1)) nucleoprotein NP gene, complete coding sequence, .

AY651491 Influenza A virus (A/Ck/Thailand/1/2004(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY651492 Influenza A virus (A/Ck/Thailand/73/2004(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY651493 Influenza A virus (A/Ck/Thailand/9.1/2004(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY651494 Influenza A virus (A/Qa/Thailand/57/2004(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY651495 Influenza A virus (A/bird/Thailand/3.1/2004(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY651496 Influenza A virus (A/Dk/Thailand/71.1/2004(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY651497 Influenza A virus (A/Gs/Thailand/79/2004(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY651502 Influenza A virus (A/Ck/Vietnam/33/2004(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY651503 Influenza A virus (A/Ck/Vietnam/35/2004(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY651504 Influenza A virus (A/Ck/Vietnam/36/2004(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY651505 Influenza A virus (A/Ck/Vietnam/37/2004(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY651506 Influenza A virus (A/Ck/Vietnam/38/2004(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY651507 Influenza A virus (A/Ck/Vietnam/39/2004(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY651508 Influenza A virus (A/Ck/Vietnam/C57/2004(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY651509 Influenza A virus (A/Dk/Vietnam/11/2004(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY651488 Influenza A virus (A/Ck/Indonesia/4/2004(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY651489 Influenza A virus (A/Ck/Indonesia/5/2004(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

AY651528 Influenza A virus (A/Peregrine/Hong Kong/D0028/2004(H5N1)) nucleocapsid protein (NP) gene, partial coding sequence.

M22573 Influenza A/Duck/Hong Kong/7/75 (H3N2), nucleoprotein (segment (seg) 5), RNA.

AY180555 Influenza A virus (A/chicken/Nanchang/3-120/2001(H3N2)) nucleoprotein (NP) gene, partial coding sequence.

AY779261 Influenza A virus (A/Turkey/North Carolina/12344/03(H3N2)) nucleoprotein (NP) gene, partial coding sequence.

AY779262 Influenza A virus (A/Turkey/Minnesota/764-2/03(H3N2)) nucleoprotein (NP) gene, partial coding sequence.

AY862655 Influenza A virus (A/chicken/Korea/S6/03(H3N2)) nucleocapsid protein (NP) gene, partial coding sequence.

AY862656 Influenza A virus (A/duck/Korea/S7/03(H3N2)) nucleocapsid protein (NP) gene, partial coding sequence.

AY862657 Influenza A virus (A/duck/Korea/S8/03(H3N2)) nucleocapsid protein (NP) gene, partial coding sequence.

AY862658 Influenza A virus (A/duck/Korea/S9/03(H3N2)) nucleocapsid protein (NP) gene, partial coding sequence.

AY862659 Influenza A virus (A/duck/Korea/S10/03(H3N2)) nucleocapsid protein (NP) gene, partial coding sequence.

AY862660 Influenza A virus (A/pigeon/Korea/S11/03(H3N2)) nucleocapsid protein (NP) gene, partial coding sequence.

D00050 Influenza A virus nucleoprotein gene, complete coding sequence.

M14921 Influenza A/Duck/NY/6750/78 (H2N2) nucleoprotein (fragment 5) RNA, complete coding sequence.

AY422026 Influenza A virus (A/duck/Hokkaido/95/01(H2N2)) nucleoprotein (NP) gene, partial coding sequence.

U49093 Influenza A virus nucleoprotein (NP) mRNA, partial coding sequence.

M22574 Influenza A/Duck/Bavaria/2/77 (H1N1), nucleoprotein (fragment 5), RNA.

M76603 Influenza A/Turkey/England/647/77(H1N1) mRNA, complete coding sequence.

M63783 Influenza A/Duck/Australia/749/80 (H1N1) nucleoprotein mRNA, complete coding sequence.

M63778 Influenza A/Turkey/Minnesota/1661/81 (H1N1) nucleoprotein mRNA, complete coding sequence.

Z26855 Influenza A virus nucleoprotein NP gene

M76609 Influenza A/Turkey/North Carolina/1790/88 (H1N1) mRNA, complete coding sequence.

Z26857 Influenza A virus nucleoprotein NP gene

AF213905 Influenza A virus (A/Duck/Italy/24/95(H1N1)) fragment 5 nucleoprotein (NP) gene, partial coding sequence.

AF213906 Influenza A virus (A/chicken/Italy/24/95(H1N1)) fragment 5 nucleoprotein (NP) gene, partial coding sequence.

AY633215 Influenza A virus (A/Mallard/Alberta/211/98(H1N1)) nucleoprotein (NP) gene, complete coding sequence.

AY180543 Influenza A virus (A/quail/Nanchang/12-340/2000(H1N1)) nucleoprotein (NP) gene, partial coding sequence.

Sequences Used in Influenza A Polymerase Basic Protein 1 (PB1) Assays

AY633218 Influenza A virus (A/Mallard/Alberta/211/98 (H1N1)) RNA-guided RNA polymerase subunit P1 (PB1) gene, partial coding sequence.

U48284 Influenza A virus polymerase (PB1) mRNA, partial coding sequence.

AY180855 Influenza A virus strain A/quail/Nanchang/12-340/2000 (H1N1) polymerase subunit PB1 (PB1) gene, partial coding sequence.

AY422038 Influenza A virus (A/duck/Hokkaido/95/01(H2N2)) polymerase subunit (PB1) gene, partial coding sequence.

M25926 Influenza A/Duck/New York/6750/78(H2N2) PB1 gene, complete coding sequence.

AY180871 Influenza A virus strain A/chicken/Nanchang/3-120/2001 (H3N2) polymerase subunit PB1 (PB1) gene, partial coding sequence.

AY779265 Influenza A virus (A/Turkey/North Carolina/12344/03 (H3N2)) polymerase basic protein 1 (PB1) gene, partial coding sequence.

AY779266 Influenza A virus (A/Turkey/Minnesota/764-2/03(H3N2)) polymerase basic protein 1 (PB1) gene, partial coding sequence.

AY862703 Influenza A virus (A/Chicken/Korea/S6/03(H3N2)) PB1(PB1) gene, partial coding sequence.

AY862704 Influenza A virus (A/duck/Korea/S7/03(H3N2)) PB1(PB1) gene, partial coding sequence.

AY862705 Influenza A virus (A/duck/Korea/S8/03(H3N2)) PB1(PB1) gene, partial coding sequence.

AY862706 Influenza A virus (A/duck/Korea/S9/03(H3N2)) PB1(PB1) gene, partial coding sequence.

AY862707 Influenza A virus (A/duck/Korea/S10/03(H3N2)) PB1(PB1) gene, partial coding sequence.

AY862708 Influenza A virus (A/pigeon/Korea/S11/03(H3N2)) PB1(PB1) gene, partial coding sequence.

AF213911 Influenza A virus (A/chicken/Italy/5945/95(H3N2)) fragment 8PB1 polymerase protein gene, partial coding sequence.

AB166860 Influenza A virus (A/Chicken/Yangkou/7/2004 (H5N1)) polymerase basic protein 1 (PB1) gene, complete coding sequence.

AB188814 Influenza A virus (A/chicken/Oita/8/2004 (H5N1)) polymerase basic protein 1 (PB1) gene, complete coding sequence.

AF398423 Influenza A virus (A/goose/Hong Kong/385.3/2000(H5N1)) polymerase (PB1) gene, partial coding sequence.

AF398424 Influenza A virus (A/goose/Hong Kong/385.5/2000(H5N1)) polymerase (PB1) gene, partial coding sequence.

AF468839 Influenza A virus (A/duck/Anyang/AVL-1/2001(H5N1)) polymerase basic protein 1 (PB1) gene, complete coding sequence.

AF509169 Influenza A virus (A/chicken/Hong Kong/FY77/01(H5N1)) polymerase (PB1) gene, partial coding sequence.

AF509170 Influenza A virus (A/chicken/Hong Kong/YU562/01(H5N1)) polymerase (PB1) gene, partial coding sequence.

AF509171 Influenza A virus (A/chicken/Hong Kong/YU563/01(H5N1)) polymerase (PB1) gene, partial coding sequence.

AF509172 Influenza A virus (A/chicken/Hong Kong/FY150/01(H5N1)) polymerase (PB1) gene, partial coding sequence.

AF509173 Influenza A virus (A/pheasant/Hong Kong/FY155/01(H5N1)) polymerase (PB1) gene, partial coding sequence.

AF509174 Influenza A virus (A/Silkie/Hong Kong/SF189/01(H5N1)) polymerase (PB1) gene, partial coding sequence.

AF509175 Influenza A virus (A/quail/Hong Kong/SF203/01(H5N1)) polymerase (PB1) gene, partial coding sequence.

AF509176 Influenza A virus (A/pigeon/Hong Kong/SF215/01(H5N1)) polymerase (PB1) gene, partial coding sequence.

AF509177 Influenza A virus (A/chicken/Hong Kong/SF219/01(H5N1)) polymerase (PB1) gene, partial coding sequence.

AF509178 Influenza A virus (A/chicken/Hong Kong/715.5/01(H5N1)) polymerase (PB1) gene, partial coding sequence.

AF509179 Influenza A virus (A/Chicken/Hong Kong/751.1/01(H5N1)) polymerase (PB1) gene, partial coding sequence.

AF509180 Influenza A virus (A/chicken/Hong Kong/822.1/01(H5N1)) polymerase (PB1) gene, partial coding sequence.

AF509181 Influenza A virus (A/Chicken/Hong Kong/829.2/01(H5N1)) polymerase (PB1) gene, partial coding sequence.

AF509182 Influenza A virus (A/Chicken/Hong Kong/830.2/01(H5N1)) polymerase (PB1) gene, partial coding sequence.

AF509183 Influenza A virus (A/chicken/Hong Kong/858.3/01(H5N1)) polymerase (PB1) gene, partial coding sequence.

AF509184 Influenza A virus (A/chicken/Hong Kong/866.3/01(H5N1)) polymerase (PB1) gene, partial coding sequence.

AF509185 Influenza A virus (A/Chicken/Hong Kong/867.1/01(H5N1)) polymerase (PB1) gene, partial coding sequence.

AF509186 Influenza A virus (A/chicken/Hong Kong/879.1/01(H5N1)) polymerase (PB1) gene, partial coding sequence.

AF509187 Influenza A virus (A/chicken/Hong Kong/873.3/01(H5N1)) polymerase (PB1) gene, partial coding sequence.

AF509188 Influenza A virus (A/chicken/Hong Kong/876.1/01(H5N1)) polymerase (PB1) gene, partial coding sequence.

AF509189 Influenza A virus (A/chicken/Hong Kong/891.1/01(H5N1)) polymerase (PB1) gene, partial coding sequence.

AF509190 Influenza A virus (A/chicken/Hong Kong/893.2/01(H5N1)) polymerase (PB1) gene, partial coding sequence.

AF509191 Influenza A virus (A/goose/Hong Kong/76.1/01(H5N1)) polymerase (PB1) gene, partial coding sequence.

AF509192 Influenza A virus (A/goose/Hong Kong/ww100/01(H5N1)) polymerase (PB1) gene, partial coding sequence.

AF509193 Influenza A virus (A/duck/Hong Kong/573.4/01(H5N1)) polymerase (PB1) gene, partial coding sequence.

AF509194 Influenza A virus (A/duck/Hong Kong/646.3/01(H5N1)) polymerase (PB1) gene, partial coding sequence.

AY035888 Influenza A virus (A/goose/Guangdong/3/97(H5N1)) polymerase basic protein 1 (PB1) gene, complete coding sequence.

AY059513 Influenza A virus (A/goose/Hong Kong/ww26/2000(H5N1)) fragment 2 polymerase (PB1) gene, partial coding sequence.

AY059514 Influenza A virus (A/goose/Hong Kong/ww28/2000(H5N1)) fragment 2 polymerase (PB1) gene, partial coding sequence.

AY059515 Influenza A virus (A/duck/Hong Kong/ww381/2000(H5N1)) fragment 2 polymerase (PB1) gene, partial coding sequence.

AY059516 Influenza A virus (A/duck/Hong Kong/ww461/2000(H5N1)) fragment 2 polymerase (PB1) gene, partial coding sequence.

AY059517 Influenza A virus (A/goose/Hong Kong/ww491/2000(H5N1)) fragment 2 polymerase (PB1) gene, partial coding sequence.

AY059518 Influenza A virus (A/duck/Hong Kong/2986.1/2000(H5N1)) fragment 2 polymerase (PB1) gene, partial coding sequence.

AY059519 Influenza A virus (A/goose/Hong Kong/3014.8/2000(H5N1)) fragment 2 polymerase (PB1) gene, partial coding sequence.

AY221575 Influenza A virus (A/chicken/Hong Kong/NT873.3/01-MB(H5N1)) polymerase basic protein 1 (PB1) gene, partial coding sequence.

AY221576 Influenza A virus (A/chicken/Hong Kong/NT873.3/01(H5N1)) polymerase basic protein 1 (PB1) gene, partial coding sequence.

AY221577 Influenza A virus (A/chicken/Hong Kong/FY150/01-MB(H5N1)) polymerase basic protein 1 (PB1) gene, partial coding sequence.

AY221578 Influenza A virus (A/chicken/Hong Kong/FY150/01(H5N1)) polymerase basic protein 1 (PB1) gene, partial coding sequence.

AY221579 Influenza A virus (A/pheasant/Hong Kong/FY155/01-MB(H5N1)) polymerase basic protein 1 (PB1) gene, partial coding sequence.

AY221580 Influenza A virus (A/pheasant/Hong Kong/FY155/01(H5N1)) polymerase basic protein 1 (PB1) gene, partial coding sequence.

AY221581 Influenza A virus (A/chicken/Hong Kong/YU822.2/01-MB(H5N1)) polymerase basic protein 1 (PB1) gene, partial coding sequence.

AY221582 Influenza A virus (A/chicken/Hong Kong/YU822.2/01(H5N1)) polymerase basic protein 1 (PB1) gene, partial coding sequence.

AY221583 Influenza A virus (A/chicken/Hong Kong/YU562/01(H5N1)) polymerase basic protein 1 (PB1) gene, partial coding sequence.

AY518366 Influenza A virus (A/Duck/China/E319-2/03(H5N1)) polymerase subunit PB1 (PB1) gene, complete coding sequence.

AY576394 Influenza A virus (A/Gs/Hong Kong/739.2/02(H5N1)) polymerase (PB1) gene, partial coding sequence.

AY576395 Influenza A virus (A/Eg/Hong Kong/757.3/02(H5N1)) polymerase (PB1) gene, partial coding sequence.

AY576396 Influenza A virus (A/G.H/Hong Kong/793.1/02(H5N1)) polymerase (PB1) gene, partial coding sequence.

AY576397 Influenza A virus (A/Dk/Hong Kong/821/02(H5N1)) polymerase (PB1) gene, complete coding sequence.

AY576398 Influenza A virus (A/Ck/Hong Kong/31.4/02(H5N1)) polymerase (PB1) gene, partial coding sequence.

AY576399 Influenza A virus (A/Ck/Hong Kong/61.9/02(H5N1)) polymerase (PB1) gene, partial coding sequence.

AY576400 Influenza A virus (A/Ck/Hong Kong/YU777/02(H5N1)) polymerase (PB1) gene, partial coding sequence.

AY576401 Influenza A virus (A/Ck/Hong Kong/96.1/02(H5N1)) polymerase (PB1) gene, partial coding sequence.

AY576402 Influenza A virus (A/Ck/Hong Kong/409.1/02(H5N1)) polymerase (PB1) gene, partial coding sequence.

AY576403 Influenza A virus (A/Ph/Hong Kong/674.15/02(H5N1)) polymerase (PB1) gene, partial coding sequence.

AY585483 Influenza A virus (A/duck/Fujian/01/2002 (H5N1)) polymerase basic protein 1 (PB1) mRNA, complete coding sequence.

AY585484 Influenza A virus (A/duck/Fujian/13/2002 (H5N1)) polymerase basic protein 1 (PB1) mRNA, complete coding sequence.

AY585485 Influenza A virus (A/duck/Fujian/17/2001(H5N1)) polymerase basic protein 1 (PB1) mRNA, complete coding sequence.

AY585486 Influenza A virus (A/duck/Fujian/19/2000(H5N1)) polymerase basic protein 1 (PB1) mRNA, complete coding sequence.

AY585487 Influenza A virus (A/duck/Guangdong/01/2001(H5N1)) polymerase basic protein 1 (PB1) mRNA, complete coding sequence.

AY585488 Influenza A virus (A/duck/Guangdong/07/2000(H5N1)) polymerase basic protein 1 (PB1) mRNA, complete coding sequence.

AY585489 Influenza A virus (A/duck/Guangdong/12/2000 (H5N1)) polymerase basic protein 1 (PB1) mRNA, complete coding sequence.

AY585490 Influenza A virus (A/duck/Guangdong/22/2002 (H5N1)) polymerase basic protein 1 (PB1) mRNA, complete coding sequence.

AY585491 Influenza A virus (A/duck/Guangdong/40/2000(H5N1)) polymerase basic protein 1 (PB1) mRNA, complete coding sequence.

AY585492 Influenza A virus (A/duck/Guangxi/07/1999 (H5N1)) polymerase basic protein 1 (PB1) mRNA, complete coding sequence.

AY585493 Influenza A virus (A/duck/Guangxi/22/2001(H5N1)) polymerase basic protein 1 (PB1) mRNA, complete coding sequence.

AY585494 Influenza A virus (A/duck/Guangxi/35/2001(H5N1)) polymerase basic protein 1 (PB1) mRNA, complete coding sequence.

AY585495 Influenza A virus (A/duck/Guangxi/50/2001(H5N1)) polymerase basic protein 1 (PB1) mRNA, complete coding sequence.

AY585496 Influenza A virus (A/duck/Guangxi/53/2002 (H5N1)) polymerase basic protein 1 (PB1) mRNA, complete coding sequence.

AY585497 Influenza A virus (A/duck/Shanghai/08/2001(H5N1)) polymerase basic protein 1 (PB1) mRNA, complete coding sequence.

AY585498 Influenza A virus (A/duck/Shanghai/13/2001(H5N1)) polymerase basic protein 1 (PB1) mRNA, complete coding sequence.

AY585499 Influenza A virus (A/duck/Shanghai/35/2002 (H5N1)) polymerase basic protein 1 (PB1) mRNA, complete coding sequence.

AY585500 Influenza A virus (A/duck/Shanghai/37/2002 (H5N1)) polymerase basic protein 1 (PB1) mRNA, complete coding sequence.

AY585501 Influenza A virus (A/duck/Shanghai/38/2001(H5N1)) polymerase basic protein 1 (PB1) mRNA, complete coding sequence.

AY585502 Influenza A virus (A/duck/Zhejiang/11/2000 (H5N1)) polymerase basic protein 1 (PB1) mRNA, complete coding sequence.

AY585503 Influenza A virus (A/duck/Zhejiang/52/2000(H5N1)) polymerase basic protein 1 (PB1) mRNA, complete coding sequence.

AY590582 Influenza A virus (A/Chicken/Nakhon Pathom/Thailand/CU-K2/2004(H5N1)) polymerase basic protein 1 (PBP1) gene, complete coding sequence.

AY609310 Influenza A virus (A/chicken/Guangdong/174/04(H5N1)) fragment 2, complete sequence.

AY651651 Influenza A virus (A/Ck/Indonesia/BL/2003(H5N1)) polymerase basic subunit 1 (PB1) gene, partial coding sequence.

AY651652 Influenza A virus (A/Dk/Indonesia/MS/2004(H5N1)) polymerase basic subunit 1 (PB1) gene, partial coding sequence.

AY651653 Influenza A virus (A/Ck/Indonesia/PA/2003(H5N1)) polymerase basic subunit 1 (PB1) gene, partial coding sequence.

AY651654 Influenza A virus (A/Ck/Indonesia/4/2004(H5N1)) polymerase basic subunit 1 (PB1) gene, partial coding sequence.

AY651655 Influenza A virus (A/Ck/Indonesia/2A/2003(H5N1)) polymerase basic subunit 1 (PB1) gene, partial coding sequence.

AY651656 Influenza A virus (A/Ck/Indonesia/5/2004(H5N1)) polymerase basic subunit 1 (PB1) gene, partial coding sequence.

AY651657 Influenza A virus (A/Ck/Thailand/1/2004(H5N1)) polymerase basic subunit 1 (PB1) gene, partial coding sequence.

AY651658 Influenza A virus (A/Ck/Thailand/73/2004(H5N1)) polymerase basic subunit 1 (PB1) gene, partial coding sequence.

AY651659 Influenza A virus (A/Ck/Thailand/9.1/2004(H5N1)) polymerase basic subunit 1 (PB1) gene, partial coding sequence.

AY651660 Influenza A virus (A/Qa/Thailand/57/2004(H5N1)) polymerase basic subunit 1 (PB1) gene, partial coding sequence.

AY651661 Influenza A virus (A/bird/Thailand/3.1/2004(H5N1)) polymerase basic subunit 1 (PB1) gene, partial coding sequence.

AY651662 Influenza A virus (A/Dk/Thailand/71.1/2004(H5N1)) polymerase basic subunit 1 (PB1) gene, partial coding sequence.

AY651663 Influenza A virus (A/Gs/Thailand/79/2004(H5N1)) polymerase basic subunit 1 (PB1) gene, partial coding sequence.

AY651668 Influenza A virus (A/Ck/Vietnam/33/2004(H5N1)) polymerase basic subunit 1 (PB1) gene, partial coding sequence.

AY651669 Influenza A virus (A/Ck/Vietnam/35/2004(H5N1)) polymerase basic subunit 1 (PB1) gene, partial coding sequence.

AY651670 Influenza A virus (A/Ck/Vietnam/36/2004(H5N1)) polymerase basic subunit 1 (PB1) gene, partial coding sequence.

AY651671 Influenza A virus (A/Ck/Vietnam/37/2004(H5N1)) polymerase basic subunit 1 (PB1) gene, partial coding sequence.

AY651672 Influenza A virus (A/Ck/Vietnam/38/2004(H5N1)) polymerase basic subunit 1 (PB1) gene, partial coding sequence.

AY651673 Influenza A virus (A/Ck/Vietnam/39/2004(H5N1)) polymerase basic subunit 1 (PB1) gene, partial coding sequence.

AY651674 Influenza A virus (A/Ck/Vietnam/C57/2004(H5N1)) polymerase basic subunit 1 (PB1) gene, partial coding sequence.

AY651675 Influenza A virus (A/Dk/Vietnam/11/2004(H5N1)) polymerase basic subunit 1 (PB1) gene, partial coding sequence.

AY651676 Influenza A virus (A/Gf/Hong Kong/38/2002 (H5N1)) polymerase basic subunit 1 (PB1) gene, partial coding sequence.

AY651677 Influenza A virus (A/Ck/Hong Kong/31.2/2002(H5N1)) polymerase basic subunit 1 gene, partial coding sequence.

AY651678 Influenza A virus (A/Ck/Hong Kong/37.4/2002 (H5N1)) polymerase basic subunit 1 (PB1) gene, partial coding sequence.

AY651679 Influenza A virus (A/SCk/Hong Kong/YU100/2002(H5N1)) polymerase basic subunit 1 (PB1) gene, partial coding sequence.

AY651680 Influenza A virus (A/Ck/Hong Kong/YU22/2002(H5N1)) polymerase basic subunit 1 (PB1) gene, partial coding sequence.

AY651681 Influenza A virus (A/Ck/Hong Kong/3176.3/2002(H5N1)) polymerase basic subunit 1 (PB1) gene, partial coding sequence.

AY651682 Influenza A virus (A/Ck/Hong Kong/3169.1/2002(H5N1)) polymerase basic subunit 1 (PB1) gene, partial coding sequence.

AY651683 Influenza A virus (A/Ck/Hong Kong/FY157/2003(H5N1)) polymerase basic subunit 1 (PB1) gene, partial coding sequence.

AY651684 Influenza A virus (A/Ck/Hong Kong/YU324/2003(H5N1)) polymerase basic subunit 1 (PB1) gene, partial coding sequence.

AY651685 Influenza A virus (A/Ck/Hong Kong/2133.1/2003(H5N1)) polymerase basic subunit 1 (PB1) gene, partial coding sequence.

AY651686 Influenza A virus (A/Ck/Hong Kong/NT93/2003(H5N1)) polymerase basic subunit 1 (PB1) gene, partial coding sequence.

AY651687 Influenza A virus (A/Ck/Hong Kong/SSP141/2003(H5N1)) polymerase basic subunit 1 (PB1) gene, partial coding sequence.

AY651688 Influenza A virus (A/Ck/Hong Kong/WF157/2003(H5N1)) polymerase basic subunit 1 (PB1) gene, partial coding sequence.

AY651689 Influenza A virus (A/Red-billed Gull/Hong Kong/12.1/2003(H5N1)) polymerase basic subunit 1 (PB1) gene, partial coding sequence.

AY651690 Influenza A virus (A/Dove/Hong Kong/862.7/2002(H5N1)) polymerase basic subunit 1 (PB1) gene, partial coding sequence.

AY651691 Influenza A virus (A/heron/Hong Kong/861.1/2002 (H5N1)) polymerase basic subunit 1 (PB1) gene, partial coding sequence.

AY651692 Influenza A virus (A/Tree Finch/Hong Kong/864/2002(H5N1)) polymerase basic subunit 1 (PB1) gene, partial coding sequence.

AY651693 Influenza A virus (A/teal/China/2978.1/2002(H5N1)) polymerase basic subunit 1 (PB1) gene, partial coding sequence.

AY651694 Influenza A virus (A/Peregrine/Hong Kong/D0028/2004(H5N1)) polymerase basic subunit 1 (PB1) gene, partial coding sequence.

AY651695 Influenza A virus (A/Dk/HN/5806/2003(H5N1)) polymerase basic subunit 1 (PB1) gene, partial coding sequence.

AY651696 Influenza A virus (A/Dk/ST/4003/2003(H5N1)) polymerase basic subunit 1 (PB1) gene, partial coding sequence.

AY651697 Influenza A virus (A/Ck/ST/4231/2003 (H5N1)) polymerase basic subunit 1 (PB1) gene, partial coding sequence.

AY651698 Influenza A virus (A/Dk/YN/6255/2003 (H5N1)) polymerase basic subunit 1 (PB1) gene, partial coding sequence.

AY651699 Influenza A virus (A/Dk/YN/6445/2003(H5N1)) polymerase basic subunit 1 (PB1) gene, partial coding sequence.

AY651700 Influenza A virus (A/Ph/ST/44/2004 (H5N1)) polymerase basic subunit 1 (PB1) gene, partial coding sequence.

AY651701 Influenza A virus (A/Dk/HN/303/2004 (H5N1)) polymerase basic subunit 1 (PB1) gene, partial coding sequence.

AY651702 Influenza A virus (A/Dk/HN/101/2004 (H5N1)) polymerase basic subunit 1 (PB1) gene, partial coding sequence.

AY651703 Influenza A virus (A/Ck/YN/374/2004 (H5N1)) polymerase basic subunit 1 (PB1) gene, partial coding sequence.

AY651704 Influenza A virus (A/Ck/YN/115/2004 (H5N1)) polymerase basic subunit 1 (PB1) gene, partial coding sequence.

AY653199 Influenza A virus (A/Chicken/Jilin/9/2004(H5N1)) fragment 2, complete sequence.

AY676025 Influenza A virus strain (A/duck/Hong Kong/821/02(H5N1)) polymerase basic protein 1 (PB1) gene, complete coding sequence.

AY676026 Influenza A virus strain (A/Egret/Hong Kong/757.2/03(H5N1)) polymerase basic protein 1 (PB1) gene, complete coding sequence.

AY676027 Influenza A virus strain (A/chicken/Korea/ES/03(H5N1)) polymerase basic protein 1 (PB1) gene, complete coding sequence.

AY676028 Influenza A virus strain (A/duck/Korea/ESD1/03(H5N1)) polymerase basic protein 1 (PB1) gene, complete coding sequence.

AY684704 Influenza A virus (A/chicken/Hubei/327/2004 (H5N1)) polymerase basic protein 1 (PB1) gene, complete coding sequence.

AY737287 Influenza A virus (A/chicken/Guangdong/191/04(H5N1)) fragment 2, complete sequence.

AY737294 Influenza A virus (A/chicken/Guangdong/178/04(H5N1)) fragment 2, complete sequence.

AY737302 Influenza A virus (A/duck/Guangdong/173/04(H5N1)) fragment 2, complete sequence.

AY770083 Influenza A virus (A/chicken/Hubei/489/2004 (H5N1)) non-functional polymerase basic protein 1 (PB1) gene, complete sequence.

AY770994 Influenza A virus (A/chicken/Ayutthaya/Thailand/CU-23/04(H5N1)) polymerase basic protein 1 gene, partial coding sequence.

AY818130 Influenza A virus (A/chicken/Vietnam/C58/04(H5N1)) polymerase protein PB1 gene, complete coding sequence.

AY818131 Influenza A virus (A/quail/Vietnam/36/04(H5N1)) polymerase protein PB1 gene, complete coding sequence.

AY856862 Influenza A virus (A/duck/Shandong/093/2004(H5N1)) fragment 2, complete sequence.

AB188822 Influenza A virus (A/chicken/Kyoto/3/2004 (H5N1)) polymerase basic protein 1 (PB1) gene, complete coding sequence.

AB189051 Influenza A virus (A/crow/Kyoto/53/2004 (H5N1)) polymerase basic protein 1 (PB1) gene, complete coding sequence, .

AB189060 Influenza A virus (A/crow/Osaka/102/2004(H5N1)) polymerase basic protein 1 (PB1) PB1 gene, complete coding sequence,.

AF046085 Influenza A virus (A/chicken/Hong Kong/220/97(H5N1)) polymerase basic protein 1 (PB1) gene, complete coding sequence.

AF098590 Influenza A virus (A/chicken/Hong Kong/258/97(H5N1)) PB1 protein (PB1) gene, partial coding sequence.

AF098591 Influenza A virus (A/chicken/Hong Kong/y388/97(H5N1)) PB1 protein (PB1) gene, partial coding sequence.

AF098592 Influenza A virus (A/chicken/Hong Kong/728/97(H5N1)) PBI protein (PB1) gene, partial coding sequence.

AF098593 Influenza A virus (A/chicken/Hong Kong/786/97(H5N1)) PB1 protein (PB1) gene, partial coding sequence.

AF098594 Influenza A virus (A/chicken/Hong Kong/915/97(H5N1)) PB1 protein (PB1) gene, partial coding sequence.

AF098595 Influenza A virus (A/duck/Hong Kong/p46/97(H5N1)) PB1 protein (PB1) gene, partial coding sequence.

AF098596 Influenza A virus (A/duck/Hong Kong/y283/97(H5N1)) PB1 protein (PB1) gene, partial coding sequence.

AF098598 Influenza A virus (A/goose/Hong Kong/w355/97(H5N1)) PB1 protein (PB1) gene, partial coding sequence.

AF144301 Influenza A virus (A/goose/Guangdong/1/96(H5N1)) polymerase (PB1) gene, complete coding sequence.

AF216716 Influenza A virus (A/environment/Hong Kong/437-4/99(H5N1)) polymerase basic protein 1 gene, complete coding sequence.

AF216724 Influenza A virus (A/environment/Hong Kong/437-6/99(H5N1)) polymerase basic protein 1 gene, complete coding sequence.

AF216732 Influenza A virus (A/environment/Hong Kong/437-8/99(H5N1)) polymerase basic protein 1 gene, complete coding sequence.

AF216740 Influenza A virus (A/Environment/Hong Kong/437-10/99(H5N1)) polymerase basic protein 1 gene, complete coding sequence.

AY303663 Influenza A virus (A/Chicken/Chile/176822/02(H7N3)) polymerase basic protein 1 gene, complete coding sequence.

AY303664 Influenza A virus (A/chicken/Chile/4957/02(H7N3)) polymerase basic protein 1 gene, partial coding sequence.

AY586435 Influenza A virus (A/Turkey/Italy/214845/02(H7N3)) PB1 gene, partial coding sequence.

AY586436 Influenza A virus (A/Turkey/Italy/220158/2002(H7N3)) PB1 gene, partial coding sequence.

AY586437 Influenza A virus (A/Duck/Italy/33/01(H7N3)) PB1 gene, partial coding sequence.

AY586438 Influenza A virus (A/Duck/Italy/43/01(H7N3)) PBI gene, partial coding sequence.

AY616765 Influenza A virus (A/chick/British Columbia/04(H7N3)) PB1 polymerase subunit (PB1) gene, complete coding sequence.

AY646084 Influenza A virus (A/Chicken/British Columbia/GSC_Human_B/04(H7N3)) polymerase basic protein 1 (PB1) gene, complete coding sequence.

AY648293 Influenza A virus (A/GSC_Chicken_B/British Columbia/04(H7N3)) PB1 polymerase subunit (PB1) gene, complete coding sequence.

AY653039 Influenza A virus (A/GSC_Chicken/British Columbia/04(H7N3)) PB1 polymerase subunit (PB1) gene, complete coding sequence.

AJ620348 Influenza A virus (A/chicken/Germany/R28/03(H7N7)) RNA polymerase PB1 gene, genomic RNA.

AY340080 Influenza A virus (A/Netherlands/124/03(H7N7)) polymerase (PB1) gene, partial coding sequence.

AY340081 Influenza A virus (A/Netherlands/126/03(H7N7)) polymerase (PB1) gene, partial coding sequence.

AY340082 Influenza A virus (A/Netherlands/127/03(H7N7)) polymerase (PB1) gene, partial coding sequence.

AY340083 Influenza A virus (A/Netherlands/219/03(H7N7)) polymerase (PB1) gene, complete coding sequence.

AY340084 Influenza A virus (A/Netherlands/033/03(H7N7)) polymerase (PB1) gene, complete coding sequence.

AY340085 Influenza A virus (A/chicken/Netherlands/1/03(H7N7)) polymerase (PB1) gene, complete coding sequence.

AB049155 Influenza A virus (A/parakeet/Chiba/1/97(H9N2)) PB1 gene for polymerase basic protein 1, complete coding sequence.

AB049156 Influenza A virus (A/parakeet/Narita/92A/98(H9N2)) polymerase basic protein 1 (PB1) gene, complete coding sequence.

AF508618 Influenza A virus (A/ostrich/South Africa/9508103/95(H9N2)) fragment 2 polymerase PB1 (PB1) gene, partial coding sequence.

AF508619 Influenza A virus (A/chicken/Pakistan/4/99(H9N2)) segment 2 polymerase PB1 (PB1) gene, partial coding sequence.

AF508620 Influenza A virus (A/chicken/Pakistan/5/99(H9N2)) fragment 2 polymerase PB1 (PB1) gene, partial coding sequence.

AF508621 Influenza A virus (A/chicken/Germany/R45/98(H9N2)) segment 2 polymerase PB1 (PB1) gene, partial coding sequence.

AF508622 Influenza A virus (A/duck/Germany/113/95(H9N2)) fragment 2 polymerase PB1 (PB1) gene, partial coding sequence.

AF508623 Influenza A virus (A/chicken/Iran/11T/99(H9N2)) segment 2 polymerase PB1 (PB1) gene, partial coding sequence.

AF508624 Influenza A virus (A/chicken/Saudi Arabia/532/99(H9N2)) segment 2 polymerase PB1 (PB1) gene, partial coding sequence.

AF508625 Influenza A virus (A/pheasant/Ireland/PV18/97(H9N2)) segment 2 polymerase PBI (PB1) gene, partial coding sequence.

AF508626 Influenza A virus (A/chicken/Korea/99029/99(H9N2)) segment 2 polymerase PB1 (PB1) gene, partial coding sequence.

AF508627 Influenza A virus (A/chicken/Beijing/8/98(H9N2)) segment 2 polymerase PB1 (PB1) gene, complete coding sequence.

AF508628 Influenza A virus (A/chicken/Guangdong/10/00(H9N2)) segment 2 polymerase PB1 (PB1) gene, partial coding sequence.

AF508629 Influenza A virus (A/chicken/Guangdong/11/97(H9N2)) segment 2 polymerase PB1 (PB1) gene, complete coding sequence.

AF508630 Influenza A virus (A/chicken/Hebei/4/98(H9N2)) segment 2 polymerase PB1 (PB1) gene, complete coding sequence.

AF508631 Influenza A virus (A/chicken/Heilongjiang/10/97(H9N2)) segment 2 polymerase PB1 (PB1) gene, partial coding sequence.

AF508632 Influenza A virus (A/chicken/Henan/62/00(H9N2)) segment 2 polymerase PB1 (PB1) gene, complete coding sequence.

AF508633 Influenza A virus (A/chicken/Ningxia/5/99(H9N2)) segment 2 polymerase PB1 (PB1) gene, complete coding sequence.

AF508634 Influenza A virus (A/Chicken/Sichuan/5/97 (H9N2)) fragment 2 polymerase PB1 (PB1) gene, partial coding sequence.

AF508635 Influenza A virus (A/chicken/Shandong/6/96(H9N2)) fragment 2PB1(PB1) gene, partial coding sequence.

AF508636 Influenza A virus (A/chicken/Shijiazhuang/2/99(H9N2)) segment 2 polymerase PB1 (PB1) gene, complete coding sequence.

AF508637 Influenza A virus (A/chicken/Shenzhen/9/97(H9N2)) fragment 2 polymerase PB1 (PB1) gene, complete coding sequence.

AF508638 Influenza A virus (A/duck/Nanjing/1/97 (H9N2)) fragment 2 polymerase PB1 (PB1) gene, complete coding sequence.

AF508639 Influenza A virus (A/quail/Shanghai/8/96(H9N2)) fragment 2 polymerase PB1 (PB1) gene, complete coding sequence.

AF523427 Influenza A virus (A/duck/Shantou/830/00(H9N2)) polymerase (PB1) gene, partial coding sequence.

AF523428 Influenza A virus (A/duck/Shantou/2102/00(H9N2)) polymerase (PB1) gene, partial coding sequence.

AF523429 Influenza A virus (A/duck/Shantou/1043/00(H9N2)) polymerase (PB1) gene, partial coding sequence.

AF523430 Influenza A virus (A/duck/Shantou/2134/00(H9N2)) polymerase (PB1) gene, partial coding sequence.

AF523431 Influenza A virus (A/wild duck/Shantou/4808/01(H9N2)) polymerase (PB1) gene, partial coding sequence.

AF523432 Influenza A virus (A/duck/Shantou/2144/00(H9N2)) polymerase (PB1) gene, partial coding sequence.

AF523433 Influenza A virus (A/duck/Shantou/2143/00(H9N2)) polymerase (PB1) gene, partial coding sequence.

AF523434 Influenza A virus (A/duck/Shantou/1796/00(H9N2)) polymerase (PB1) gene, partial coding sequence.

AF523435 Influenza A virus (A/duck/Shantou/2088/01(H9N2)) polymerase (PB1) gene, partial coding sequence.

AF523436 Influenza A virus (A/duck/Shantou/1881/00(H9N2)) polymerase (PB1) gene, partial coding sequence.

AF523437 Influenza A virus (A/duck/Hong Kong/366/78(H9N2)) polymerase (PB1) gene, partial coding sequence.

AF523438 Influenza A virus (A/duck/Hong Kong/552/79(H9N2)) polymerase (PB1) gene, partial coding sequence.

AF523439 Influenza A virus (A/duck/Hong Kong/86/76(H9N2)) polymerase (PB1) gene, partial coding sequence.

AF523440 Influenza A virus (A/duck/Hong Kong/289/78(H9N2)) polymerase (PB1) gene, partial coding sequence.

AF523441 Influenza A virus (A/duck/Hong Kong/610/79(H9N2)) polymerase (PB1) gene, partial coding sequence.

AF523442 Influenza A virus (A/duck/Shantou/1605/01(H9N2)) polymerase (PB1) gene, partial coding sequence.

AF523443 Influenza A virus (A/duck/Shantou/1042/00(H9N2)) polymerase (PB1) gene, partial coding sequence.

AF536659 Influenza A virus (A/chicken/Beijing/1/95(H9N2)) PB1 gene, partial coding sequence.

AF536660 Influenza A virus (A/chicken/Beijing/2/97(H9N2)) PB1 gene, partial coding sequence.

AF536661 Influenza A virus (A/chicken/Beijing/3/99(H9N2)) PB1 gene, partial coding sequence.

AF536662 Influenza A virus (A/chicken/Guangdong/97(H9N2)) PB1 gene, partial coding sequence.

AF536663 Influenza A virus (A/chicken/Hebei/1/96(H9N2)) PB1 gene, partial coding sequence.

AF536664 Influenza A virus (A/chicken/Hebei/2/98(H9N2)) PB1 gene, partial coding sequence.

AF536665 Influenza A virus (A/chicken/Hebei/3/98(H9N2)) PBI gene, partial coding sequence.

AF536666 Influenza A virus (A/chicken/Henan/98(H9N2)) PB1 gene, partial coding sequence.

AF536667 Influenza A virus (A/chicken/Liaoning/99(H9N2)) PBI gene, partial coding sequence.

AF536668 Influenza A virus (A/Chicken/Shandong/98(H9N2)) PB1 gene, partial coding sequence.

AJ291396 Influenza A virus (A/chicken/Pakistan/2/99(H9N2)) polymerase PB1 (PB1) gene, genomic RNA.

AJ427862 Influenza A virus (A/quail/Hong Kong/FY298/00(H9N2)) partial PB1 polymerase protein pb1 gene, genomic RNA

AY180840 Influenza A virus strain A/pigeon/Nanchang/7-058/2000 (H9N2) polymerase subunit PB1 (PB1) gene, partial coding sequence.

AY180843 Influenza A virus strain A/quail/Nanchang/2-0460/2000 (H9N2) polymerase subunit PB1 (PB1) gene, partial coding sequence.

AY180844 Influenza A virus strain A/pigeon/Nanchang/2-0461/2000 (H9N2) polymerase subunit PB1 (PB1) gene, partial coding sequence.

AY180851 Influenza A virus strain A/pigeon/Nanchang/11-145/2000 (H9N2) polymerase subunit PB1 (PB1) gene, partial coding sequence.

AY180852 Influenza A virus strain A/duck/Nanchang/11-197/2000 (H9N2) polymerase subunit PB1 (PB1) gene, partial coding sequence.

AY180854 Influenza A virus strain A/duck/Nanchang/11-290/2000 (H9N2) polymerase subunit PB1 (PB1) gene, partial coding sequence.

AY180856 Influenza A virus strain A/duck/Nanchang/1-0070/2000 (H9N2) polymerase subunit PB1 (PB1) gene, partial coding sequence.

AY180866 Influenza A virus strain A/duck/Nanchang/7-092/2000 (H9N2) polymerase subunit PB1 (PB1) gene, partial coding sequence.

AY180867 Influenza A virus strain A/chicken/Nanchang/1-0016/2000 (H9N2) polymerase subunit PB1 (PB1) gene, partial coding sequence.

AY180873 Influenza A virus strain A/chicken/Nanchang/4-010/2000 (H9N2) polymerase subunit PB1 (PB1) gene, partial coding sequence.

AY180874 Influenza A virus strain A/chicken/Nanchang/4-301/2001 (H9N2) polymerase subunit PB1 (PB1) gene, partial coding sequence.

AY180875 Influenza A virus strain A/chicken/Nanchang/4-361/2001 (H9N2) polymerase subunit PB1 (PB1) gene, partial coding sequence.

AY180892 Influenza A virus strain A/quail/Nanchang/4-040/2000 (H9N2) polymerase subunit PB1 (PB1) gene, partial coding sequence.

AY180897 Influenza A virus strain A/Wild duck/Nanchang/2-0480/2000 (H9N2) polymerase subunit PB1 (PB1) gene, partial coding sequence.

AY180900 Influenza A virus strain A/duck/Nanchang/10-389/2000 (H9N2) polymerase subunit PB1 (PB1) gene, partial coding sequence.

AY180901 Influenza A virus strain A/duck/Nanchang/11-392/2000 (H9N2) polymerase subunit PB1 (PB1) gene, partial coding sequence.

AY253751 Influenza A virus (A/chicken/Shanghai/F/98(H9N2)) polymerase basic protein 1 (PB1) gene, complete coding sequence.

AY307947 Influenza A virus (A/chicken/Beijing/1/00(H9N2)) polymerase subunit (PB1) gene, partial coding sequence.

AY307948 Influenza A virus (A/chicken/Hebei/1/01(H9N2)) polymerase subunit (PB1) gene, partial coding sequence.

AY633170 Influenza A virus (A/Mallard/Alberta/17/91(H9N2)) RNA-guided RNA polymerase subunit P1 (PB1) gene, partial coding sequence.

AY633282 Influenza A virus (A/Mallard/Alberta/321/88(H9N2)) RNA-guided RNA polymerase subunit P1 (PB1) gene, partial coding sequence.

AY633298 Influenza A virus (A/Mallard/Alberta/11/91(H9N2)) RNA-guided RNA polymerase subunit P1 (PB1) gene, partial coding sequence.

AY664774 Influenza A virus (A/chicken/Hong Kong/CSW153/03(H9N2)) polymerase basic protein 1 (PB1) gene, partial coding sequence.

AY664775 Influenza A virus (A/chicken/Hong Kong/AP45/03(H9N2)) polymerase basic protein 1 (PB1) gene, partial coding sequence.

AY664776 Influenza A virus (A/chicken/Hong Kong/BD90/03(H9N2)) polymerase basic protein 1 (PB1) gene, partial coding sequence.

AY664777 Influenza A virus (A/chicken/Hong Kong/CSW291/03(H9N2)) polymerase basic protein 1 (PB1) gene, partial coding sequence.

AY664778 Influenza A virus (A/chicken/Hong Kong/CSW304/03(H9N2)) polymerase basic protein 1 (PB1) gene, partial coding sequence.

AY664779 Influenza A virus (A/chicken/Hong Kong/FY23/03(H9N2)) polymerase basic protein 1 (PB1) gene, partial coding sequence.

AY664780 Influenza A virus (A/Grizzly chicken/Hong Kong/NT101/03(H9N2)) polymerase basic protein 1 (PB1) gene, partial coding sequence.

AY664781 Influenza A virus (A/chicken/Hong Kong/NT142/03(H9N2)) polymerase basic protein 1 (PB1) gene, partial coding sequence.

AY664782 Influenza A virus (A/chicken/Hong Kong/SF1/03(H9N2)) polymerase basic protein 1 (PB1) gene, partial coding sequence.

AY664783 Influenza A virus (A/chicken/Hong Kong/SSP101/03(H9N2)) polymerase basic protein 1 (PB1) gene, partial coding sequence.

AY664784 Influenza A virus (A/chicken/Hong Kong/TP38/03(H9N2)) polymerase basic protein 1 (PB1) gene, partial coding sequence.

AY664785 Influenza A virus (A/chicken/Hong Kong/WF126/03(H9N2)) polymerase basic protein 1 (PB1) gene, partial coding sequence.

AY664786 Influenza A virus (A/pigeon/Hong Kong/WF53/03(H9N2)) polymerase basic protein 1 (PB1) gene, partial coding sequence.

AY664787 Influenza A virus (A/pheasant/Hong Kong/WF54/03(H9N2)) polymerase basic protein 1 (PB1) gene, partial coding sequence.

AY664788 Influenza A virus (A/Grizzly Chicken/Hong Kong/NT184/03(H9N2)) polymerase basic protein 1 (PB1) gene, partial coding sequence.

AY664789 Influenza A virus (A/chicken/Hong Kong/WF120/03(H9N2)) polymerase basic protein 1 (PB1) gene, partial coding sequence.

AY664790 Influenza A virus (A/chicken/Hong Kong/NT366/03(H9N2)) polymerase basic protein 1 (PB1) gene, partial coding sequence.

AY664791 Influenza A virus (A/chicken/Hong Kong/YU427/03(H9N2)) polymerase basic protein 1 (PB1) gene, partial coding sequence.

AY800239 Influenza A virus (A/chicken/Korea/S1/2003(H9N2)) polymerase basic protein 1 (PB1) gene, partial coding sequence.

AY862694 Influenza A virus (A/silky chicken/Korea/S3/03(H9N2)) PB1 (PB1) gene, partial coding sequence.

AY862695 Influenza A virus (A/Chicken/Korea/S4/03(H9N2)) PB1(PB1) gene, partial coding sequence.

AY862696 Influenza A virus (A/Chicken/Korea/S5/03(H9N2)) PB1(PB1) gene, partial coding sequence.

AY862697 Influenza A virus (A/Chicken/Korea/S12/03(H9N2)) PB1(PB1) gene, partial coding sequence.

AY862698 Influenza A virus (A/duck/Korea/S13/03(H9N2)) PB1(PB1) gene, partial coding sequence.

AY862699 Influenza A virus (A/pigeon/Korea/S14/03(H9N2)) PB1(PB1) gene, partial coding sequence.

AY862700 Influenza A virus (A/Chicken/Korea/S15/03(H9N2)) PB1(PB1) gene, partial coding sequence.

AY862701 Influenza A virus (A/Chicken/Korea/S16/03(H9N2)) PBI(PB1) gene, partial coding sequence.

AY862702 Influenza A virus (A/Chicken/Korea/S18/03(H9N2)) PB1(PB1) gene, partial coding sequence.

AF156416 Influenza A virus (A/chicken/Hong Kong/G9/97(H9N2)) fragment 2 PB1 polymerase subunit (PB1) gene, partial coding sequence.

AF156417 Influenza A virus (A/chicken/Hong Kong/G23/99(H9N2)) fragment 2 PB1 polymerase subunit (PB1) gene, partial coding sequence.

AF156418 Influenza A virus (A/pigeon/Hong Kong/Y233/97(H9N2)) fragment 2 PB1 polymerase subunit (PB1) gene, partial coding sequence.

AF156419 Influenza A virus (A/duck/Hong Kong/Y280/97(H9N2)) fragment 2 PB1 polymerase subunit (PB1) gene, partial coding sequence.

AF156420 Influenza A virus (A/duck/Hong Kong/Y439/97(H9N2)) fragment 2 PB1 polymerase subunit (PB1) gene, partial coding sequence.

AF156421 Influenza A virus (A/quail/Hong Kong/G1/97(H9N2)) fragment 2 PB1 polymerase subunit (PB1) gene, partial coding sequence.

AF156422 Influenza A virus (A/chicken/Hong Kong/739/94(H9N2)) fragment 2 PB1 polymerase subunit (PB1) gene, partial coding sequence.

AF156423 Influenza A virus (A/chicken/Beijing/1/94(H9N2)) fragment 2 PB1 polymerase subunit (PB1) gene, partial coding sequence.

AF156424 Influenza A virus (A/quail/Hong Kong/AF157/92(H9N2)) fragment 2 PB1 polymerase subunit (PB1) gene, partial coding sequence.

AF156425 Influenza A virus (A/chicken/Korea/38349-p96323/96(H9N2)) fragment 2 PB1 polymerase subunit (PB1) gene, partial coding sequence.

AF156426 Influenza A virus (A/chicken/Korea/25232-96006/96(H9N2)) fragment 2 PB1 polymerase subunit (PB1) gene, partial coding sequence.

AF156427 Influenza A virus (A/shorebird/Delaware/9/96(H9N2)) fragment 2 PB1 polymerase subunit (PB1) gene, partial coding sequence.

AF156428 Influenza A virus (A/quail/Arkansas/29209-1/93(H9N2)) fragment 2 PB1 polymerase subunit (PB1) gene, partial coding sequence.

AF156429 Influenza A virus (A/Turkey/California/189/66(H9N2)) fragment 2 PB1 polymerase subunit (PB1) gene, partial coding sequence.

AF222632 Influenza A virus (A/quail/Hong Kong/A17/99(H9N2)) fragment 2 polymerase 1 (PB1) gene, partial coding sequence.

AF222633 Influenza A virus (A/pigeon/Hong Kong/FY6/99(H9N2)) fragment 2 polymerase 1 (PB1) gene, partial coding sequence.

AF222634 Influenza A virus (A/chicken/Hong Kong/NT16/99(H9N2)) fragment 2 polymerase 1 (PB1) gene, partial coding sequence.

AF222635 Influenza A virus (A/quail/Hong Kong/SSP10/99(H9N2)) fragment 2 polymerase 1 (PB1) gene, partial coding sequence.

AF222636 Influenza A virus (A/pheasant/Hong Kong/SSP11/99(H9N2)) fragment 2 polymerase 1 (PB1) gene, partial coding sequence.

AF222637 Influenza A virus (A/chicken/Hong Kong/FY20/99(H9N2)) fragment 2 polymerase 1 (PB1) gene, partial coding sequence.

AF222638 Influenza A virus (A/chicken/Hong Kong/KC12/99(H9N2)) segment 2 polymerase 1 (PB1) gene, partial coding sequence.

AF222639 Influenza A virus (A/quail/Hong Kong/NT2899(H9N2)) segment 2 polymerase 1 (PB1) gene, partial coding sequence.

AF222640 Influenza A virus (A/chicken/Hong Kong/SF2/99(H9N2)) segment 2 polymerase 1 (PB1) gene, partial coding sequence.

AF222641 Influenza A virus (A/Silky chick/Hong Kong/SF44/99(H9N2)) fragment 2 polymerase 1 (PB1) gene, partial coding sequence.

Sequences Used in Influenza A Polymerase Basic Protein 2 (PB2) Assays

gi|49356919|AY633219| Influenza A virus (A/Mallard/Alberta/211/98 (H1N1)) polymerase basic protein 2 (PB2) gene, partial coding sequence.

gi|27466107|AY180748| Influenza A virus strain A/quail/Nanchang/12-340/2000 (H1N1) polymerase subunit PB2 (PB2) gene, partial coding sequence.

gi|45272173|AY422042| Influenza A virus (A/duck/Hokkaido/95/01(H2N2)) polymerase subunit (PB2) gene, partial coding sequence.

gi|18091825|AF213910| Influenza A virus (A/chicken/Italy/5945/95(H3N2)) fragment 1PB2 polymerase protein gene, partial coding sequence.

gi|27466133|AY180761| Influenza A virus strain A/chicken/Nanchang/3-120/2001 (H3N2) polymerase subunit PB2 (PB2) gene, partial coding sequence.

gi|56160002|AY779267| Influenza A virus (A/Turkey/North Carolina/12344/03 (H3N2)) polymerase basic protein 2 (PB2) gene, partial coding sequence.

gi|56160004|AY779268| Influenza A virus (A/Turkey/Minnesota/764-2/03 (H3N2)) polymerase basic protein 2 (PB2) gene, partial coding sequence.

gi|58429704|AY862719| Influenza A virus (A/Chicken/Korea/S6/03(H3N2)) PB2 (PB2) gene, partial coding sequence.

gi|58429706|AY862720| Influenza A virus (A/duck/Korea/S7/03(H3N2)) PB2 (PB2) gene, partial coding sequence.

gi|58429708|AY862721| Influenza A virus (A/duck/Korea/S8/03(H3N2)) PB2 (PB2) gene, partial coding sequence.

gi|58429710|AY862722| Influenza A virus (A/duck/Korea/S9/03(H3N2)) PB2 (PB2) gene, partial coding sequence.

gi|58429712|AY862723| Influenza A virus (A/duck/Korea/S10/03(H3N2)) PB2 (PB2) gene, partial coding sequence.

gi|58429714|AY862724| Influenza A virus (A/pigeon/Korea/S11/03(H3N2)) PB2 (PB2) gene, partial coding sequence.

gi|5805276|AF144300| Influenza A virus (A/goose/Guangdong/1/96(H5N1)) polymerase (PB2) gene, complete coding sequence.

gi|3335416|AF046086| Influenza A virus (A/chicken/Hong Kong/220/97 (H5N1)) polymerase basic protein 2 (PB2) gene, partial coding sequence.

gi|6048841|AF098577| Influenza A virus (A/Chicken/Hong Kong/258/97(H5N1)) PB2 protein (PB2) gene, partial coding sequence.

gi|6048843|AF098578| Influenza A virus (A/chicken/Hong Kong/y388/97(H5N1)) PB2protein(PB2) gene, partial coding sequence.

gi|6048845|AF098579| Influenza A virus (A/Chicken/Hong Kong/728/97(H5N1)) PB2 protein (PB2) gene, partial coding sequence.

gi|6048847|AF098580| Influenza A virus (A/chicken/Hong Kong/786/97(H5N1)) PB2 protein (PB2) gene, partial coding sequence.

gi|6048849|AF098581| Influenza A virus (A/Chicken/Hong Kong/915/97(H5N1)) PB2 protein (PB2) gene, partial coding sequence.

gi|6048851|AF098582| Influenza A virus (A/duck/Hong Kong/p46/97(H5N1)) PB2 protein (PB2) gene, partial coding sequence.

gi|6048853|AF098583| Influenza A virus (A/duck/Hong Kong/y283/97(H5N1)) PB2 protein (PB2) gene, partial coding sequence.

gi|6048855|AF098584| Influenza A virus (A/goose/Hong Kong/w355/97(H5N1)) PB2 protein (PB2) gene, partial coding sequence.

gi|14860983|AY038798| Influenza A virus (A/goose/Guangdong/3/1997(H5N1)) PB2 protein (PB2) gene, complete coding sequence.

gi|47156244|AY585513| Influenza A virus (A/duck/Guangxi/07/1999 (H5N1)) polymerase basic protein 2 (PB2) mRNA, complete coding sequence.

gi|9863884|AF216717| Influenza A virus (A/environment/Hong Kong/437-4/99(H5N1)) polymerase basic protein 2 gene, partial coding sequence.

gi|9863903|AF216725| Influenza A virus (A/environment/Hong Kong/437-6/99(H5N1)) polymerase basic protein 2 gene, partial coding sequence.

gi|9863921|AF216733| Influenza A virus (A/environment/Hong Kong/437-8/99(H5N1)) polymerase basic protein 2 gene, partial coding sequence.

gi|9863939|AF216741| Influenza A virus (A/environment/Hong Kong/437-10/99(H5N1)) polymerase basic protein 2 gene, partial coding sequence.

gi|47156264|AY585523| Influenza A virus (A/duck/Zhejiang/11/2000 (H5N1)) polymerase basic protein 2 (PB2) mRNA, complete coding sequence.

gi|47156266|AY585524| Influenza A virus (A/duck/Zhejiang/52/2000 (H5N1)) polymerase basic protein 2 (PB2) mRNA, complete coding sequence.

gi|47156232|AY585507| Influenza A virus (A/duck/Fujian/19/2000 (H5N1)) polymerase basic protein 2 (PB2) mRNA, complete coding sequence.

gi|47156236|AY585509| Influenza A virus (A/duck/Guangdong/07/2000(H5N1)) polymerase basic protein 2 (PB2) mRNA, complete coding sequence.

gi|47156238|AY585510| Influenza A virus (A/duck/Guangdong/12/2000 (H5N1)) polymerase basic protein 2 (PB2) mRNA, complete coding sequence.

gi|47156242|AY585512| Influenza A virus (A/duck/Guangdong/40/2000 (H5N1)) polymerase basic protein 2 (PB2) mRNA, complete coding sequence.

gi|19697849|AY059520| Influenza A virus (A/goose/Hong Kong/ww26/2000(H5N1)) fragment 1 polymerase (PB2) gene, partial coding sequence.

gi|19697851|AY059521| Influenza A virus (A/goose/Hong Kong/ww28/2000(H5N1)) fragment 1 polymerase (PB2) gene, partial coding sequence.

gi|19697853|AY059522| Influenza A virus (A/duck/Hong Kong/ww381/2000 (H5N1)) fragment 1 polymerase (PB2) gene, partial coding sequence.

gi|19697855|AY059523| Influenza A virus (A/duck/Hong Kong/ww461/2000(H5N1)) fragment 1 polymerase (PB2) gene, partial coding sequence.

gi|19697857|AY059524| Influenza A virus (A/goose/Hong Kong/ww491/2000(H5N1)) fragment 1 polymerase (PB2) gene, partial coding sequence.

gi|19697859|AY059525| Influenza A virus (A/duck/Hong Kong/2986.1/2000(H5N1)) fragment 1 polymerase (PB2) gene, partial coding sequence.

gi|19697867|AY059529| Influenza A virus (A/goose/Hong Kong/3014.8/2000 (H5N1)) fragment 1 polymerase (PB2) gene, partial coding sequence.

gi|18092181|AF398425| Influenza A virus (A/goose/Hong Kong/385.3/2000 (H5N1)) polymerase (PB2) gene, partial coding sequence.

gi|18092183|AF398426| Influenza A virus (A/goose/Hong Kong/385.5/2000 (H5N1)) polymerase (PB2) gene, partial coding sequence.

gi|21359665|AF468840| Influenza A virus (A/duck/Anyang/AVL-1/2001 (H5N1)) polymerase basic protein 2 (PB2) gene, partial coding sequence.

gi|28849606|AF509143| Influenza A virus (A/chicken/Hong Kong/FY77/01(H5N1)) polymerase (PB2) gene, partial coding sequence.

gi|28849608|AF509144| Influenza A virus (A/chicken/Hong Kong/YU562/01(H5N1)) polymerase (PB2) gene, partial coding sequence.

gi|28849610|AF509145| Influenza A virus (A/chicken/Hong Kong/YU563/01(H5N1)) polymerase (PB2) gene, partial coding sequence.

gi|28849612|AF509146| Influenza A virus (A/chicken/Hong Kong/FY150/01(H5N1)) polymerase (PB2) gene, partial coding sequence.

gi|28849614|AF509147| Influenza A virus (A/pheasant/Hong Kong/FY155/01(H5N1)) polymerase (PB2) gene, partial coding sequence.

gi|28849616|AF509148| Influenza A virus (A/Zhusiji/Hong Kong/SF189/01(H5N1)) polymerase (PB2) gene, partial coding sequence.

gi|28849618|AF509149| Influenza A virus (A/quail/Hong Kong/SF203/01(H5N1)) polymerase (PB2) gene, partial coding sequence.

gi|28849620|AF509150| Influenza A virus (A/pigeon/Hong Kong/SF215/01(H5N1)) polymerase (PB2) gene, partial coding sequence.

gi|28849622|AF509151| Influenza A virus (A/chicken/Hong Kong/SF219/01(H5N1)) polymerase (PB2) gene, partial coding sequence.

gi|28849624|AF509152| Influenza A virus (A/chicken/Hong Kong/715.5/01(H5N1)) polymerase (PB2) gene, partial coding sequence.

gi|28849626|AF509153| Influenza A virus (A/chicken/Hong Kong/751.1/01(H5N1)) polymerase (PB2) gene, partial coding sequence.

gi|28849628|AF509154| Influenza A virus (A/chicken/Hong Kong/822.1/01(H5N1)) polymerase (PB2) gene, partial coding sequence.

gi|28849630|AF509155| Influenza A virus (A/chicken/Hong Kong/829.2/01(H5N1)) polymerase (PB2) gene, partial coding sequence.

gi|28849632|AF509156| Influenza A virus (A/chicken/Hong Kong/830.2/01(H5N1)) polymerase (PB2) gene, partial coding sequence.

gi|28849634|AF509157| Influenza A virus (A/chicken/Hong Kong/858.3/01(H5N1)) polymerase (PB2) gene, partial coding sequence.

gi|28849636|AF509158| Influenza A virus (A/chicken/Hong Kong/866.3/01(H5N1)) polymerase (PB2) gene, partial coding sequence.

gi|28849638|AF509159| Influenza A virus (A/chicken/Hong Kong/867.1/01(H5N1)) polymerase (PB2) gene, partial coding sequence.

gi|28849640|AF509160| Influenza A virus (A/chicken/Hong Kong/879.1/01(H5N1)) polymerase (PB2) gene, partial coding sequence.

gi|28849642|AF509161| Influenza A virus (A/chicken/Hong Kong/873.3/01(H5N1)) polymerase (PB2) gene, partial coding sequence.

gi|28849644|AF509162| Influenza A virus (A/chicken/Hong Kong/876.1/01(H5N1)) polymerase (PB2) gene, partial coding sequence.

gi|28849646|AF509163| Influenza A virus (A/chicken/Hong Kong/891.1/01 (H5N1)) polymerase (PB2) gene, partial coding sequence.

gi|28849648|AF509164| Influenza A virus (A/chicken/Hong Kong/893.2/01(H5N1)) polymerase (PB2) gene, partial coding sequence.

gi|28849650|AF509165| Influenza A virus (A/goose/Hong Kong/76.1/01(H5N1)) polymerase (PB2) gene, partial coding sequence.

gi|28849652|AF509166| Influenza A virus (A/goose/Hong Kong/ww100/01(H5N1)) polymerase (PB2) gene, partial coding sequence.

gi|28849654|AF509167| Influenza A virus (A/duck/Hong Kong/573.4/01(H5N1)) polymerase (PB2) gene, partial coding sequence.

gi|28849656|AF509168| Influenza A virus (A/duck/Hong Kong/646.3/01(H5N1)) polymerase (PB2) gene, partial coding sequence.

gi|28823262|AY221584| Influenza A virus (A/chicken/Hong Kong/NT873.3/01-MB(H5N1)) polymerase basic protein 2 (PB2) gene, partial coding sequence.

gi|28823443|AY221585| Influenza A virus (A/chicken/Hong Kong/NT873.3/01(H5N1)) polymerase basic protein 2 (PB2) gene, partial coding sequence.

gi|28823612|AY221586| Influenza A virus (A/chicken/Hong Kong/FY150/01-MB(H5N1)) polymerase basic protein 2 (PB2) gene, partial coding sequence.

gi|28823783|AY221587| Influenza A virus (A/chicken/Hong Kong/FY150/01(H5N1)) polymerase basic protein 2 (PB2) gene, partial coding sequence.

gi|28823961|AY221588| Influenza A virus (A/pheasant/Hong Kong/FY155/01-MB(H5N1)) polymerase basic protein 2 (PB2) gene, partial coding sequence.

gi|28824143|AY221589| Influenza A virus (A/pheasant/Hong Kong/FY155/01(H5N1)) polymerase basic protein 2 (PB2) gene, partial coding sequence.

gi|28824334|AY221590| Influenza A virus (A/chicken/Hong Kong/YU822.2/01-MB(H5N1)) polymerase basic protein 2 (PB2) gene, partial coding sequence.

gi|28824502|AY221591| Influenza A virus (A/chicken/Hong Kong/YU822.2/01(H5N1)) polymerase basic protein 2 (PB2) gene, partial coding sequence.

gi|28824684|AY221592| Influenza A virus (A/chicken/Hong Kong/YU562/01(H5N1)) polymerase basic protein 2 (PB2) gene, partial coding sequence.

gi|47156230|AY585506| Influenza A virus (A/duck/Fujian/17/2001 (H5N1)) polymerase basic protein 2 (PB2) mRNA, complete coding sequence.

gi|47156234|AY585508| Influenza A virus (A/duck/Guangdong/01/2001 (H5N1)) polymerase basic protein 2 (PB2) mRNA, complete coding sequence.

gi|47156246|AY585514| Influenza A virus (A/duck/Guangxi/22/2001 (H5N1)) polymerase basic protein 2 (PB2) mRNA, complete coding sequence.

gi|47156248|AY585515| Influenza A virus (A/duck/Guangxi/35/2001 (H5N1)) polymerase basic protein 2 (PB2) mRNA, complete coding sequence.

gi|47156250|AY585516| Influenza A virus (A/duck/Guangxi/50/2001 (H5N1)) polymerase basic protein 2 (PB2) mRNA, complete coding sequence.

gi|47156254|AY585518| Influenza A virus (A/duck/Shanghai/08/2001(H5N1)) polymerase basic protein 2 (PB2) mRNA, complete coding sequence.

gi|47156256|AY585519| Influenza A virus (A/duck/Shanghai/13/2001(H5N1)) polymerase basic protein 2 (PB2) mRNA, complete coding sequence.

gi|47156262|AY585522| Influenza A virus (A/duck/Shanghai/38/2001 (H5N1)) polymerase basic protein 2 (PB2) mRNA, complete coding sequence.

gi|47156258|AY585520| Influenza A virus (A/duck/Shanghai/35/2002 (H5N1)) polymerase basic protein 2 (PB2) mRNA, complete coding sequence.

gi|47156260|AY585521| Influenza A virus (A/duck/Shanghai/37/2002 (H5N1)) polymerase basic protein 2 (PB2) mRNA, complete coding sequence.

gi|47156252|AY585517| Influenza A virus (A/duck/Guangxi/53/2002 (H5N1)) polymerase basic protein 2 (PB2) mRNA, complete coding sequence.

gi|47156240|AY585511| Influenza A virus (A/duck/Guangdong/22/2002 (H5N1)) polymerase basic protein 2 (PB2) mRNA, complete coding sequence.

gi|47834791|AY576382| Influenza A virus (A/Gs/Hong Kong/739.2/02 (H5N1)) polymerase (PB2) gene, partial coding sequence.

gi|47834793|AY576383| Influenza A virus (A/Eg/Hong Kong/757.3/02 (H5N1)) polymerase (PB2) gene, partial coding sequence.

gi|47834795|AY576384| Influenza A virus (A/G.H/Hong Kong/793.1/02(H5N1)) polymerase (PB2) gene, partial coding sequence.

gi|47834797|AY576385| Influenza A virus (A/Dk/Hong Kong/821/02(H5N1)) polymerase (PB2) gene, partial coding sequence.

gi|47834799|AY576386| Influenza A virus (A/Ck/Hong Kong/31.4/02(H5N1)) polymerase (PB2) gene, partial coding sequence.

gi|47834801|AY576387| Influenza A virus (A/Ck/Hong Kong/61.9/02(H5N1)) polymerase (PB2) gene, partial coding sequence.

gi|47834803|AY576388| Influenza A virus (A/Ck/Hong Kong/YU777/02(H5N1)) polymerase (PB2) gene, partial coding sequence.

gi|47834805|AY576389| Influenza A virus (A/Ck/Hong Kong/96.1/02(H5N1)) polymerase (PB2) gene, partial coding sequence.

gi|47834807|AY576390| Influenza A virus (A/Ck/Hong Kong/409.1/02(H5N1)) polymerase (PB2) gene, partial coding sequence.

gi|47834809|AY576391| Influenza A virus (A/Ph/Hong Kong/sv674.15/02(H5N1)) polymerase (PB2) gene, partial coding sequence.

gi|47156226|AY585504| Influenza A virus (A/duck/Fujian/01/2002 (H5N1)) polymerase basic protein 2 (PB2) mRNA, complete coding sequence.

gi|47156228|AY585505| Influenza A virus (A/duck/Fujian/13/2002 (H5N1)) polymerase basic protein 2 (PB2) mRNA, complete coding sequence.

gi|50296597|AY651744| Influenza A virus (A/heron/Hong Kong/861.1/2002 (H5N1)) polymerase basic subunit 2 (PB2) gene, partial coding sequence.

gi|50296599|AY651745| Influenza A virus (A/Dove/Hong Kong/862.7/2002 (H5N1)) polymerase basic subunit 2 (PB2) gene, partial coding sequence.

gi|50296601|AY651746| Influenza A virus (A/Tree Finch/Hong Kong/864/2002 (H5N1)) polymerase basic subunit 2 (PB2) gene, partial coding sequence.

gi|50296603|AY651747| Influenza A virus (A/teal/China/2978.1/2002 (H5N1)) polymerase basic subunit 2 (PB2) gene, partial coding sequence.

gi|56548879|AY676021| Influenza A virus (A/duck/Hong Kong/821/02(H5N1)) polymerase basic protein 2 (PB2) gene, complete coding sequence.

gi|50296569|AY651730| Influenza A virus (A/Gf/Hong Kong/38/2002 (H5N1)) polymerase basic subunit 2 (PB2) gene, partial coding sequence.

gi|50296571|AY651731| Influenza A virus (A/Ck/Hong Kong/31.2/2002 (H5N1)) polymerase basic subunit 2 (PB2) gene, partial coding sequence.

gi|50296573|AY651732| Influenza A virus (A/Ck/Hong Kong/37.4/2002 (H5N1)) polymerase basic subunit 2 (PB2) gene, partial coding sequence.

gi|50296575|AY651733| Influenza A virus (A/SCk/Hong Kong/YU100/2002 (H5N1)) polymerase basic subunit 2 (PB2) gene, partial coding sequence.

gi|50296577|AY651734| Influenza A virus (A/Ck/Hong Kong/YU22/2002 (H5N1)) polymerase basic subunit 2 (PB2) gene, partial coding sequence.

gi|50296579|AY651735| Influenza A virus (A/Ck/Hong Kong/3176.3/2002 (H5N1)) polymerase basic subunit 2 (PB2) gene, partial coding sequence.

gi|50296581|AY651736| Influenza A virus (A/Ck/Hong Kong/3169.1/2002 (H5N1)) polymerase basic subunit 2 (PB2) gene, partial coding sequence.

gi|50296583|AY651737| Influenza A virus (A/Ck/Hong Kong/FY157/2003 (H5N1)) polymerase basic subunit 2 (PB2) gene, partial coding sequence.

gi|50296585|AY651738| Influenza A virus (A/Ck/Hong Kong/YU324/2003 (H5N1)) polymerase basic subunit 2 (PB2) gene, partial coding sequence.

gi|50296587|AY651739| Influenza A virus (A/Ck/Hong Kong/2133.1/2003 (H5N1)) polymerase basic subunit 2 (PB2) gene, partial coding sequence.

gi|50296589|AY651740| Influenza A virus (A/Ck/Hong Kong/NT93/2003 (H5N1)) polymerase basic subunit 2 (PB2) gene, partial coding sequence.

gi|50296591|AY651741| Influenza A virus (A/Ck/Hong Kong/SSP141/2003 (H5N1)) polymerase basic subunit 2 (PB2) gene, partial coding sequence.

gi|50296593|AY651742| Influenza A virus (A/Ck/Hong Kong/WF157/2003 (H5N1)) polymerase basic subunit 2 (PB2) gene, partial coding sequence.

gi|50296595|AY651743| Influenza A virus (A/Red-billed Gull/Hong Kong/12.1/2003 (H5N1)) polymerase basic subunit 2 (PB2) gene, partial coding sequence.

gi|50296607|AY651749| Influenza A virus (A/Dk/HN/5806/2003 (H5N1)) polymerase basic subunit 2 (PB2) gene, partial coding sequence.

gi|50296609|AY651750| Influenza A virus (A/Dk/ST/4003/2003 (H5N1)) polymerase basic subunit 2 (PB2) gene, partial coding sequence.

gi|50296611|AY651751| Influenza A virus (A/Ck/ST/4231/2003 (H5N1)) polymerase basic subunit 2 (PB2) gene, partial coding sequence.

gi|50296613|AY651752| Influenza A virus (A/Dk/YN/6255/2003 (H5N1)) polymerase basic subunit 2 (PB2) gene, partial coding sequence.

gi|50296615|AY651753| Influenza A virus (A/Dk/YN/6445/2003 (H5N1)) polymerase basic subunit 2 (PB2) gene, partial coding sequence.

gi|50296529|AY651710| Influenza A virus (A/Ck/Indonesia/2A/2003 (H5N1)) polymerase basic subunit 2 (PB2) gene, partial coding sequence.

gi|56548881|AY676022| Influenza A virus (A/Egret/Hong Kong/757.2/03(H5N1)) polymerase basic protein 2 (PB2) gene, complete coding sequence.

gi|56548883|AY676023| Influenza A virus (A/Chicken/Korea/ES/03(H5N1)) polymerase basic protein 2 (PB2) gene, complete coding sequence.

gi|56548885|AY676024| Influenza A virus (A/Duck/Korea/ESD1/03(H5N1)) polymerase basic protein 2 (PB2) gene, complete coding sequence.

gi|50296519|AY651705| Influenza A virus (A/Ck/Indonesia/PA/2003(H5N1)) polymerase basic subunit 2 (PB2) gene, complete coding sequence.

gi|50296523|AY651707| Influenza A virus (A/Ck/Indonesia/BL/2003(H5N1)) polymerase basic subunit 2 (PB2) gene, complete coding sequence.

gi|41207501|AY518367| Influenza A virus (A/duck/China/E319-2/03(H5N1)) polymerase subunit PB2 (PB2) gene, complete coding sequence.

gi|45359369|AY550147| Influenza A virus (A/chicken/Nakhon Pathom/Thailand/CU-K2/04(H5N1)) polymerase basic protein 2 (PB2) gene, partial coding sequence.

gi|50296525|AY651708| Influenza A virus (A/Ck/Indonesia/5/2004 (H5N1)) polymerase basic subunit 2 (PB2) gene, partial coding sequence.

gi|50296527|AY651709| Influenza A virus (A/Ck/Indonesia/4/2004 (H5N1)) polymerase basic subunit 2 (PB2) gene, partial coding sequence.

gi|50296521|AY651706| Influenza A virus (A/Dk/Indonesia/MS/2004(H5N1)) polymerase basic subunit 2 (PB2) gene, complete coding sequence.

gi|51094103|AY590581| Influenza A virus (A/chicken/Nakhon Pathom/Thailand/CU-K2/2004(H5N1)) polymerase basic protein 2 (PBP2) gene, partial coding sequence.

gi|47716766|AY609309| Influenza A virus (A/chicken/Guangdong/174/04(H5N1)) fragment 1, complete sequence.

gi|58531082|AB166859| Influenza A virus (A/Chicken/Yamaguchi/7/2004(H5N1)) PB2 gene for polymerase basic protein 2, complete coding sequence.

gi|58531114|AB188813| Influenza A virus (A/chicken/Oita/8/2004(H5N1)) PB2 gene of polymerase basic protein 2, complete coding sequence.

gi|50956621|AY684703| Influenza A virus (A/chicken/Hubei/327/2004 (H5N1)) polymerase basic protein 2 (PB2) gene, complete coding sequence.

gi|57915957|AY737286| Influenza A virus (A/chicken/Guangdong/191/04(H5N1)) fragment 1, complete sequence.

gi|57916006|AY737293| Influenza A virus (A/chicken/Guangdong/178/04(H5N1)) fragment 1, complete sequence.

gi|57916060|AY737301| Influenza A virus (A/duck/Guangdong/173/04(H5N1)) fragment 1, complete sequence.

gi|55233237|AY770084| Influenza A virus (A/chicken/Hubei/489/2004 (H5N1)) polymerase basic protein 2 (PB2) gene, complete coding sequence.

gi|54873461|AY770993| Influenza A virus (A/chick/Ayutthaya/Thailand/CU-23/04(H5N1)) polymerase basic protein 2 gene, partial coding sequence.

gi|58618421|AY818127| Influenza A virus (A/chicken/Vietnam/C58/04(H5N1)) polymerase protein PB2 gene, complete coding sequence.

gi|58618423|AY818128| Influenza A virus (A/quail/Vietnam/36/04(H5N1)) polymerase protein PB2 gene, complete coding sequence.

gi|58374183|AY856861| Influenza A virus (A/duck/Shandong/093/2004(H5N1)) fragment 1, complete sequence.

gi|58531132|AB188821| Influenza A virus (A/Chicken/Kyoto/3/2004(H5N1)) PB2 gene of polymerase basic protein 2, complete coding sequence.

gi|58531150|AB189050| Influenza A virus (A/crow/Kyoto/53/2004(H5N1)) PB2 gene of polymerase basic protein 2, complete coding sequence, .

gi|58531168|AB189058| Influenza A virus (A/crow/Osaka/102/2004(H5N1)) PB2 gene of polymerase basic protein 2, complete coding sequence.

gi|50296605|AY651748| Influenza A virus (A/Peregrine/Hong Kong/D0028/2004(H5N1)) polymerase basic subunit 2 (PB2) gene, complete coding sequence.

gi|50296531|AY651711| Influenza A virus (A/Ck/Thailand/1/2004 (H5N1)) polymerase basic subunit 2 (PB2) gene, partial coding sequence.

gi|50296533|AY651712| Influenza A virus (A/Ck/Thailand/73/2004 (H5N1)) polymerase basic subunit 2 (PB2) gene, partial coding sequence.

gi|50296535|AY651713| Influenza A virus (A/Ck/Thailand/9.1/2004 (H5N1)) polymerase basic subunit 2 (PB2) gene, complete coding sequence.

gi|50296537|AY651714| Influenza A virus (A/Qa/Thailand/57/2004 (H5N1)) polymerase basic subunit 2 (PB2) gene, complete coding sequence.

gi|50296539|AY651715| Influenza A virus (A/bird/Thailand/3.1/2004 (H5N1)) polymerase basic subunit 2 (PB2) gene, complete coding sequence.

gi|50296541|AY651716| Influenza A virus (A/Dk/Thailand/71.1/2004 (H5N1)) polymerase basic subunit 2 (PB2) gene, complete coding sequence.

gi|50296543|AY651717| Influenza A virus (A/Gs/Thailand/79/2004 (H5N1)) polymerase basic subunit 2 (PB2) gene, partial coding sequence.

gi|50296553|AY651722| Influenza A virus (A/Ck/Vietnam/33/2004 (H5N1)) polymerase basic subunit 2 (PB2) gene, complete coding sequence.

gi|50296555|AY651723| Influenza A virus (A/Ck/Vietnam/35/2004 (H5N1)) polymerase basic subunit 2 (PB2) gene, complete coding sequence.

gi|50296557|AY651724| Influenza A virus (A/Ck/Vietnam/36/2004 (H5N1)) polymerase basic subunit 2 (PB2) gene, complete coding sequence.

gi|50296559|AY651725| Influenza A virus (A/Ck/Vietnam/37/2004 (H5N1)) polymerase basic subunit 2 (PB2) gene, complete coding sequence.

gi|50296561|AY651726| Influenza A virus (A/Ck/Vietnam/38/2004 (H5N1)) polymerase basic subunit 2 (PB2) gene, complete coding sequence.

gi|50296563|AY651727| Influenza A virus (A/Ck/Vietnam/39/2004 (H5N1)) polymerase basic subunit 2 (PB2) gene, complete coding sequence.

gi|50296565|AY651728| Influenza A virus (A/Ck/Vietnam/C57/2004 (H5N1)) polymerase basic subunit 2 (PB2) gene, complete coding sequence.

gi|50296567|AY651729| Influenza A virus (A/Dk/Vietnam/11/2004 (H5N1)) polymerase basic subunit 2 (PB2) gene, complete coding sequence.

gi|50296617|AY651754| Influenza A virus (A/Ck/YN/374/2004 (H5N1)) polymerase basic subunit 2 (PB2) gene, partial coding sequence.

gi|50296619|AY651755| Influenza A virus (A/Ck/YN/115/2004 (H5N1)) polymerase basic subunit 2 (PB2) gene, partial coding sequence.

gi|50296621|AY651756| Influenza A virus (A/Ph/ST/44/2004 (H5N1)) polymerase basic subunit 2 (PB2) gene, partial coding sequence.

gi|50296623|AY651757| Influenza A virus (A/Dk/HN/303/2004 (H5N1)) polymerase basic subunit 2 (PB2) gene, partial coding sequence.

gi|50296625|AY651758| Influenza A virus (A/Dk/HN/101/2004 (H5N1)) polymerase basic subunit 2 (PB2) gene, partial coding sequence.

gi|50365712|AY653193| Influenza A virus (A/chicken/Jilin/9/2004(H5N1)) fragment 1, complete sequence.

gi|47680940|AY586445| Influenza A virus (A/Duck/Italy/43/01(H7N3)) PB2 gene, partial coding sequence.

gi|47680930|AY586440| Influenza A virus (A/Duck/Italy/33/01(H7N3)) PB2 gene, partial coding sequence.

gi|47680932|AY586441| Influenza A virus (A/Turkey/Italy/220158/2002(H7N3)) PB2 gene, partial coding sequence.

gi|45124743|AJ627485| Influenza A virus (A/Turkey/Italy/214845/2002 (H7N3)) RNA polymerase PB2 gene, genomic RNA.

gi|45124767|AJ627496| Influenza A virus (A/Turkey/Italy/220158/2002 (H7N3)) RNA polymerase PB2 gene, genomic RNA.

gi|34597782|AY303665| Influenza A virus (A/Chicken/Chile/176822/02(H7N3)) polymerase basic protein 2 gene, partial coding sequence.

gi|34597784|AY303666| Influenza A virus (A/Chicken/Chile/4957/02(H7N3)) polymerase basic protein 2 gene, partial coding sequence.

gi|47680928|AY586439| Influenza A virus (A/Turkey/Italy/214845/02(H7N3)) PB2 gene, partial coding sequence.

gi|47834374|AY616766| Influenza A virus (A/chick/British Columbia/04(H7N3)) PB2 polymerase subunit (PB2) gene, complete coding sequence.

gi|50542651|AY646085| Influenza A virus (A/Chicken/British Columbia/GSC_Human_B/04(H7N3)) polymerase basic protein 2 (PB2) gene, complete coding sequence.

gi|50083053|AY648294| Influenza A virus (A/GSC_Chick_B/British Columbia/04(H7N3)) PB2 polymerase subunit (PB2) gene, complete coding sequence.

gi|50059194|AY650276| Influenza A virus (A/GSC_Chicken/British Columbia/04(H7N3)) PB2 polymerase subunit (PB2) gene, complete coding sequence.

gi|60700|X58691| Influenza A virus (A/FPV/Waterfish/27(H7N7) PB2 gene m-cap binding protein, genomic RNA

gi|325001|M38291| Influenza A/FPV/Weybridge polymerase basic protein 2 (PB2) (fragment 3) gene, complete coding sequence.

gi|9988661|AF268120| Influenza A virus (A/RedKnot/Delaware/259/94(H7N7)) polymerase protein PB2 gene, partial coding sequence.

gi|40732893|AJ620347| Influenza A virus ((A/Chicken/Germany/R28/03(H7N7))A/Chicken/Germany/R28/03(H7N7)) RNA polymerase PB2 gene, genomic RNA.

gi|37813157|AY342410| Influenza A virus (A/Netherlands/124/03(H7N7)) polymerase protein 2 gene, partial coding sequence.

gi|37813159|AY342411| Influenza A virus (A/Netherlands/126/03(H7N7)) polymerase protein 2 gene, partial coding sequence.

gi|37813161|AY342412| Influenza A virus (A/Netherlands/127/03(H7N7)) polymerase protein 2 gene, partial coding sequence.

gi|37813163|AY342413| Influenza A virus (A/Netherlands/219/03(H7N7)) polymerase protein 2 gene, partial coding sequence.

gi|37813165|AY342414| Influenza A virus (A/chick/Netherlands/1/03(H7N7)) polymerase protein 2 gene, partial coding sequence.

gi|5732354|AF156443| Influenza A virus (A/Turkey/California/189/66(H9N2)) fragment 1PB2 polymerase subunit (PB2) gene, partial coding sequence.

gi|31339587|AF523469| Influenza A virus (A/duck/Hong Kong/86/76(H9N2)) polymerase protein (PB2) gene, partial coding sequence.

gi|31339583|AF523467| Influenza A virus (A/duck/Hong Kong/366/78 (H9N2)) polymerase (PB2) gene, partial coding sequence.

gi|31339605|AF523478| Influenza A virus (A/duck/Hong Kong/289/78(H9N2)) polymerase (PB2) gene, partial coding sequence.

gi|31339585|AF523468| Influenza A virus (A/duck/Hong Kong/552/79(H9N2)) polymerase (PB2) gene, partial coding sequence.

gi|31339589|AF523470| Influenza A virus (A/duck/Hong Kong/610/79 (H9N2)) polymerase (PB2) gene, partial coding sequence.

gi|49356935|AY633283| Influenza A virus (A/Mallard/Alberta/321/88 (H9N2)) polymerase basic protein 2 (PB2) gene, partial coding sequence.

gi|49356939|AY633299| Influenza A virus (A/Mallard/Alberta/11/91 (H9N2)) polymerase basic protein 2 (PB2) gene, partial coding sequence.

gi|49356907|AY633171| Influenza A virus (A/Mallard/Alberta/17/91 (H9N2)) polymerase basic protein 2 (PB2) gene, partial coding sequence.

gi|5732342|AF156437| Influenza A virus (A/quail/Hong Kong/AF157/92(H9N2)) fragment 1 PB2 polymerase subunit (PB2) gene, partial coding sequence.

gi|5732352|AF156442| Influenza A virus (A/quail/Arkansas/29209-1/93(H9N2)) fragment 1 PB2 polymerase subunit (PB2) gene, partial coding sequence.

gi|5732340|AF156436| Influenza A virus (A/chicken/Hong Kong/739/94(H9N2)) fragment 1 PB2 polymerase subunit (PB2) gene, partial coding sequence.

gi|5732344|AF156438| Influenza A virus (A/chicken/Beijing/1/94(H9N2)) fragment 1 PB2 polymerase subunit (PB2) gene, partial coding sequence.

gi|22759060|AF536679| Influenza A virus (A/chicken/Beijing/1/95(H9N2)) PB2 gene, partial coding sequence.

gi|33318110|AF508640| Influenza A virus (A/ostrich/South Africa/9508103/95 (H9N2)) segment 1 polymerase PB2 (PB2) gene, complete coding sequence.

gi|33318118|AF508644| Influenza A virus (A/duck/Germany/113/95(H9N2)) fragment 1 polymerase PB2 (PB2) gene, partial coding sequence.

gi|33318144|AF508657| Influenza A virus (A/chicken/Shandong/6/96(H9N2)) fragment 1 polymerase PB2 (PB2) gene, partial coding sequence.

gi|33318152|AF508661| Influenza A virus (A/quail/Shanghai/8/96 (H9N2)) fragment 1 polymerase PB2 (PB2) gene, partial coding sequence.

gi|5732346|AF156439| Influenza A virus (A/chicken/Korea/38349-p96323/96(H9N2)) fragment 1 PB2 polymerase subunit (PB2) gene, partial coding sequence.

gi|5732348|AF156440| Influenza A virus (A/chicken/Korea/25232-96006/96(H9N2)) fragment 1 PB2 polymerase subunit (PB2) gene, partial coding sequence.

gi|5732350|AF156441| Influenza A virus (A/Shorebird/Delaware/9/96(H9N2)) fragment 1 PB2 polymerase subunit (PB2) gene, partial coding sequence.

gi|22759068|AF536683| Influenza A virus (A/chicken/Hebei/1/96(H9N2)) PB2 gene, partial coding sequence.

gi|5732328|AF156430| Influenza A virus (A/Chicken/Hong Kong/G9/97(H9N2)) fragment 1 PB2 polymerase subunit (PB2) gene, complete coding sequence.

gi|5732330|AF156431| Influenza A virus (A/chicken/Hong Kong/G23/97(H9N2)) fragment 1 PB2 polymerase subunit (PB2) gene, partial coding sequence.

gi|5732332|AF156432| Influenza A virus (A/pigeon/Hong Kong/Y233/97(H9N2)) fragment 1 PB2 polymerase subunit (PB2) gene, partial coding sequence.

gi|5732334|AF156433| Influenza A virus (A/duck/Hong Kong/Y280/97(H9N2)) fragment 1 PB2 polymerase subunit (PB2) gene, partial coding sequence.

gi|5732336|AF156434| Influenza A virus (A/duck/Hong Kong/Y439/97(H9N2)) fragment 1 PB2 polymerase subunit (PB2) gene, partial coding sequence.

gi|5732338|AF156435| Influenza A virus (A/quail/Hong Kong/G1/97(H9N2)) fragment 1 PB2 polymerase subunit (PB2) gene, partial coding sequence.

gi|33318148|AF508659| Influenza A virus (A/chicken/Shenzhen/9/97 (H9N2)) fragment 1 polymerase PB2 (PB2) gene, partial coding sequence.

gi|33318150|AF508660| Influenza A virus (A/duck/Nanjing/1/97 (H9N2)) fragment 1 polymerase PB2 (PB2) gene, partial coding sequence.

gi|33318124|AF508647| Influenza A virus (A/pheasant/Ireland/PV18/97 (H9N2)) fragment 1 polymerase PB2 (PB2) gene, partial coding sequence.

gi|13383266|AB049153| Influenza A virus (A/parakeet/Chiba/1/97(H9N2)) polymerase basic protein 2 PB2 gene, complete coding sequence.

gi|33318132|AF508651| Influenza A virus (A/chicken/Guangdong/11/97(H9N2)) fragment 1 polymerase PB2 (PB2) gene, partial coding sequence.

gi|33318136|AF508653| Influenza A virus (A/chicken/Heilongjiang/10/97(H9N2)) fragment 1 polymerase PB2 (PB2) gene, partial coding sequence.

gi|22759062|AF536680| Influenza A virus (A/chicken/Beijing/2/97(H9N2)) PB2 gene, partial coding sequence.

gi|33318142|AF508656| Influenza A virus (A/Chicken/Sichuan/5/97(H9N2)) fragment 1 polymerase PB2 (PB2) gene, partial coding sequence.

gi|22759066|AF536682| Influenza A virus (A/chicken/Guangdong/97(H9N2)) PB2 gene, partial coding sequence.

gi|22759078|AF536688| Influenza A virus (A/chicken/Shandong/98(H9N2)) PB2 gene, partial coding sequence.

gi|33318116|AF508643| Influenza A virus (A/chicken/Germany/R45/98(H9N2)) fragment 1 polymerase PB2 (PB2) gene, partial coding sequence.

gi|33318134|AF508652| Influenza A virus (A/chicken/Hebei/4/98(H9N2)) fragment 1 polymerase PB2 (PB2) gene, partial coding sequence.

gi|33318128|AF508649| Influenza A virus (A/chicken/Beijing/8/98 (H9N2)) fragment 1 polymerase PB2 (PB2) gene, complete coding sequence.

gi|13383268|AB049154| Influenza A virus (A/parakeet/Narita/92A/98(H9N2)) polymerase basic protein 2 PB2 gene, complete coding sequence.

gi|22759070|AF536684| Influenza A virus (A/chicken/Hebei/2/98(H9N2)) PB2 gene, partial coding sequence.

gi|22759072|AF536685| Influenza A virus (A/chicken/Hebei/3/98(H9N2)) PB2 gene, partial coding sequence.

gi|22759074|AF536686| Influenza A virus (A/chicken/Henan/98(H9N2)) PB2 gene, partial coding sequence.

gi|30025722|AY253750| Influenza A virus (A/chicken/Shanghai/F/98(H9N2)) RNA polymerase (PB2) gene, complete coding sequence.

gi|12060631|AF222622| Influenza A virus (A/quail/Hong Kong/A17/99(H9N2)) fragment 1 polymerase 2 (PB2) gene, partial coding sequence.

gi|12060633|AF222623| Influenza A virus (A/pigeon/Hong Kong/FY6/99(H9N2)) fragment 1 polymerase 2 (PB2) gene, partial coding sequence.

gi|12060635|AF222624| Influenza A virus (A/chicken/Hong Kong/NT16/99(H9N2)) segment 1 polymerase 2 (PB2) gene, partial coding sequence.

gi|12060637|AF222625| Influenza A virus (A/quail/Hong Kong/SSP10/99(H9N2)) segment 1 polymerase 2 (PB2) gene, partial coding sequence.

gi|12060639|AF222626| Influenza A virus (A/pheasant/Hong Kong/SSP11/99(H9N2)) segment 1 polymerase 2 (PB2) gene, partial coding sequence.

gi|12060641|AF222627| Influenza A virus (A/chicken/Hong Kong/FY20/99(H9N2)) fragment 1 polymerase 2 (PB2) gene, partial coding sequence.

gi|12060643|AF222628| Influenza A virus (A/chicken/Hong Kong/KC12/99(H9N2)) fragment 1 polymerase 2 (PB2) gene, partial coding sequence.

gi|12060645|AF222629| Influenza A virus (A/quail/Hong Kong/NT28/99(H9N2)) segment 1 polymerase 2 (PB2) gene, partial coding sequence.

gi|12060647|AF222630| Influenza A virus (A/chicken/Hong Kong/SF2/99(H9N2)) segment 1 polymerase 2 (PB2) gene, partial coding sequence.

gi|12060649|AF222631| Influenza A virus (A/Silky chick/Hong Kong/SF44/99(H9N2)) fragment 1 polymerase 2 (PB2) gene, partial coding sequence.

gi|22759076|AF536687| Influenza A virus (A/chicken/Liaoning/99(H9N2)) PB2 gene, partial coding sequence.

gi|33318112|AF508641| Influenza A virus (A/Chicken/Pakistan/4/99(H9N2)) fragment 1 polymerase PB2 (PB2) gene, complete coding sequence.

gi|33318114|AF508642| Influenza A virus (A/Chicken/Pakistan/5/99(H9N2)) fragment 1 polymerase PB2 (PB2) gene, complete coding sequence.

gi|33318126|AF508648| Influenza A virus (A/chicken/Korea/99029/99(H9N2)) segment 1 polymerase PB2 (PB2) gene, partial coding sequence.

gi|33318120|AF508645| Influenza A virus (A/chick/Iran/11T/99(H9N2)) segment 1 polymerase PB2 (PB2) gene, complete coding sequence.

gi|33318122|AF508646| Influenza A virus (A/Chicken/Saudi Arabia/532/99(H9N2)) fragment 1 polymerase PB2 (PB2) gene, partial coding sequence.

gi|33318140|AF508655| Influenza A virus (A/chicken/Ningxia/5/99 (H9N2)) fragment 1 polymerase PB2 (PB2) gene, partial coding sequence.

gi|33318146|AF508658| Influenza A virus (A/chicken/Shijiazhuang/2/99 (H9N2)) fragment 1 polymerase PB2 (PB2) gene, partial coding sequence.

gi|12038893|AJ291395| Influenza A virus (A/chick/Pakistan/2/99(H9N2)) polymerase PB2 gene, genomic RNA.

gi|22759064|AF536681| Influenza A virus (A/chicken/Beijing/3/99(H9N2)) PB2 gene, partial coding sequence.

gi|31339607|AF523479| Influenza A virus (A/duck/Shantou/1881/00(H9N2)) polymerase (PB2) gene, partial coding sequence.

gi|31339601|AF523476| Influenza A virus (A/duck/Shantou/830/00(H9N2)) polymerase (PB2) gene, partial coding sequence.

gi|31339603|AF523477| Influenza A virus (A/duck/Shantou/1796/00 (H9N2)) polymerase (PB2) gene, partial coding sequence.

gi|18496117|AJ427861| Influenza A virus (A/quail/Hong Kong/FY298/00(H9N2)) PB2 polymerase part pb2 gene, genomic RNA

gi|27466041|AY180715| Influenza A virus strain A/wild duck/Nanchang/2-0480/2000 (H9N2) polymerase subunit PB2 (PB2) gene, partial coding sequence.

gi|27466043|AY180716| Influenza A virus strain A/pigeon/Nanchang/2-0461/2000 (H9N2) polymerase subunit PB2 (PB2) gene, partial coding sequence.

gi|27466045|AY180717| Influenza A virus strain A/duck/Nanchang/1-0070/2000 (H9N2) polymerase subunit PB2 (PB2) gene, partial coding sequence.

gi|27466055|AY180722| Influenza A virus strain A/duck/Nanchang/10-389/2000 (H9N2) polymerase subunit PB2 (PB2) gene, partial coding sequence.

gi|27466057|AY180723| Influenza A virus strain A/pigeon/Nanchang/7-058/2000 (H9N2) polymerase subunit PB2 (PB2) gene, partial coding sequence.

gi|27466067|AY180728| Influenza A virus strain A/quail/Nanchang/2-0460/2000 (H9N2) polymerase subunit PB2 (PB2) gene, partial coding sequence.

gi|27466085|AY180737| Influenza A virus strain A/pigeon/Nanchang/11-145/2000 (H9N2) polymerase subunit PB2 (PB2) gene, partial coding sequence.

gi|27466087|AY180738| Influenza A virus strain A/duck/Nanchang/11-197/2000 (H9N2) polymerase subunit PB2 (PB2) gene, partial coding sequence.

gi|27466091|AY180740| Influenza A virus strain A/duck/Nanchang/11-290/2000 (H9N2) polymerase subunit PB2 (PB2) gene, partial coding sequence.

gi|27466093|AY180741| Influenza A virus strain A/duck/Nanchang/11-392/2000 (H9N2) polymerase subunit PB2 (PB2) gene, partial coding sequence.

gi|31339575|AF523463| Influenza A virus (A/duck/Shantou/2134/00 (H9N2)) polymerase (PB2) gene, partial coding sequence.

gi|31339579|AF523465| Influenza A virus (A/duck/Shantou/1043/00(H9N2)) polymerase (PB2) gene, partial coding sequence.

gi|31339581|AF523466| Influenza A virus (A/duck/Shantou/1042/00 (H9N2)) polymerase (PB2) gene, partial coding sequence.

gi|31339593|AF523472| Influenza A virus (A/duck/Shantou/2102/00(H9N2)) polymerase (PB2) gene, partial coding sequence.

gi|31339595|AF523473| Influenza A virus (A/duck/Shantou/2144/00(H9N2)) polymerase (PB2) gene, partial coding sequence.

gi|31339597|AF523474| Influenza A virus (A/duck/Shantou/2143/00(H9N2)) polymerase (PB2) gene, partial coding sequence.

gi|33318138|AF508654| Influenza A virus (A/chicken/Henan/62/00(H9N2)) fragment 1 polymerase PB2 (PB2) gene, partial coding sequence.

gi|33318130|AF508650| Influenza A virus (A/chicken/Guangdong/10/00(H9N2)) fragment 1 polymerase PB2 (PB2) gene, partial coding sequence.

gi|27466121|AY180755| Influenza A virus strain A/duck/Nanchang/7-092/2000 (H9N2) polymerase subunit PB2 (PB2) gene, partial coding sequence.

gi|27466141|AY180765| Influenza A virus strain A/chicken/Nanchang/4-010/2000 (H9N2) polymerase subunit PB2 (PB2) gene, partial coding sequence.

gi|27466157|AY180773| Influenza A virus strain A/quail/Nanchang/4-040/2000 (H9N2) polymerase subunit PB2 (PB2) gene, partial coding sequence.

gi|27466159|AY180774| Influenza A virus strain A/chicken/Nanchang/1-0016/2000 (H9N2) polymerase subunit PB2 (PB2) gene, partial coding sequence.

gi|55469788|AY768575| Influenza A virus (A/chicken/Korea/SNU0028/00(H9N2)) polymerase basic subunit 2 (PB2) gene, partial coding sequence.

gi|55469790|AY768576| Influenza A virus (A/chicken/Korea/SNU0037/00(H9N2)) polymerase basic subunit 2 (PB2) gene, partial coding sequence.

gi|55469792|AY768577| Influenza A virus (A/chicken/Korea/SNU0073/00(H9N2)) polymerase basic subunit 2 (PB2) gene, partial coding sequence.

gi|55469794|AY768578| Influenza A virus (A/chicken/Korea/SNU0091/00(H9N2)) polymerase basic subunit 2 (PB2) gene, partial coding sequence.

gi|55469796|AY768579| Influenza A virus (A/chicken/Korea/SNU0140/00(H9N2)) polymerase basic subunit 2 (PB2) gene, partial coding sequence.

gi|55469798|AY768580| Influenza A virus (A/chicken/Korea/SNU0146/00(H9N2)) polymerase basic subunit 2 (PB2) gene, partial coding sequence.

gi|55469800|AY768581| Influenza A virus (A/chicken/Korea/SNU1035C/00(H9N2)) polymerase basic subunit 2 (PB2) gene, partial coding sequence.

gi|27466143|AY180766| Influenza A virus strain A/chicken/Nanchang/4-301/2001 (H9N2) polymerase subunit PB2 (PB2) gene, partial coding sequence.

gi|31339599|AF523475| Influenza A virus (A/duck/Shantou/2088/01(H9N2)) polymerase (PB2) gene, partial coding sequence.

gi|31339591|AF523471| Influenza A virus (A/duck/Shantou/1605/01(H9N2)) polymerase (PB2) gene, partial coding sequence.

gi|31339577|AF523464| Influenza A virus (A/wild duck/Shantou/4808/01(H9N2)) polymerase (PB2) gene, partial coding sequence.

gi|27466097|AY180743| Influenza A virus strain A/chicken/Nanchang/4-361/2001 (H9N2) polymerase subunit PB2 (PB2) gene, partial coding sequence.

gi|54398631|AY664792| Influenza A virus (A/chicken/Hong Kong/CSW153/03(H9N2)) polymerase basic protein 2 (PB2) gene, partial coding sequence.

gi|54398633|AY664793| Influenza A virus (A/chicken/Hong Kong/AP45/03(H9N2)) polymerase basic protein 2 (PB2) gene, partial coding sequence.

gi|54398635|AY664794| Influenza A virus (A/chicken/Hong Kong/BD90/03(H9N2)) polymerase basic protein 2 (PB2) gene, partial coding sequence.

gi|54398637|AY664795| Influenza A virus (A/chicken/Hong Kong/CSW291/03(H9N2)) non-functional polymerase basic protein 2 (PB2) gene, partial sequence.

gi|54398638|AY664796| Influenza A virus (A/chicken/Hong Kong/CSW304/03(H9N2)) non-functional polymerase basic protein 2 (PB2) gene, partial sequence.

gi|54398639|AY664797| Influenza A virus (A/chicken/Hong Kong/FY23/03(H9N2)) non-functional polymerase basic protein 2 (PB2) gene, partial sequence.

gi|54398640|AY664798| Polymerase basic protein 2 (PB2) gene of influenza A virus (A/Grizzly chicken/Hong Kong/NT101/03(H9N2)), partial coding sequence.

gi|54398642|AY664799| Influenza A virus (A/chicken/Hong Kong/NT142/03(H9N2)) polymerase basic protein 2 (PB2) gene, partial coding sequence.

gi|54398644|AY664800| Influenza A virus (A/chicken/Hong Kong/SF1/03(H9N2)) non-functional polymerase basic protein 2 (PB2) gene, partial sequence.

gi|54398645|AY664801| Influenza A virus (A/chicken/Hong Kong/SSP101/03(H9N2)) non-functional polymerase basic protein 2 (PB2) gene, partial sequence.

gi|54398646|AY664802| Influenza A virus (A/chicken/Hong Kong/TP38/03(H9N2)) polymerase basic protein 2 (PB2) gene, partial coding sequence.

gi|54398648|AY664803| Influenza A virus (A/chicken/Hong Kong/WF126/03(H9N2)) non-functional polymerase basic protein 2 (PB2) gene, partial sequence.

gi|54398649|AY664804| Influenza A virus (A/pigeon/Hong Kong/WF53/03(H9N2)) non-functional polymerase basic protein 2 (PB2) gene, partial sequence.

gi|54398650|AY664805| Influenza A virus (A/pheasant/Hong Kong/WF54/03(H9N2)) non-functional polymerase basic protein 2 (PB2) gene, partial sequence.

gi|54398651|AY664806| Polymerase basic protein 2 (PB2) gene of influenza A virus (A/Grizzly chicken/Hong Kong/NT184/03(H9N2)), partial coding sequence.

gi|54398653|AY664807| Influenza A virus (A/chicken/Hong Kong/WF120/03(H9N2)) non-functional polymerase basic protein 2 (PB2) gene, partial sequence.

gi|54398654|AY664808| Influenza A virus (A/chicken/Hong Kong/NT366/03(H9N2)) polymerase basic protein 2 (PB2) gene, partial coding sequence.

gi|54398656|AY664809| Influenza A virus (A/chicken/Hong Kong/SSP418/03(H9N2)) polymerase basic protein 2 (PB2) gene, partial coding sequence.

gi|54398658|AY664810| Influenza A virus (A/chicken/Hong Kong/YU427/03(H9N2)) polymerase basic protein 2 (PB2) gene, partial coding sequence.

gi|55793686|AY800240| Influenza A virus (A/chicken/Korea/S1/2003 (H9N2)) polymerase basic protein 2 (PB2) gene, partial coding sequence.

gi|58429686|AY862710| Influenza A virus (A/Silkie/Korea/S3/03(H9N2)) PB2 (PB2) gene, partial coding sequence.

gi|58429688|AY862711| Influenza A virus (A/Chicken/Korea/S4/03(H9N2)) PB2 (PB2) gene, partial coding sequence.

gi|58429690|AY862712| Influenza A virus (A/Chicken/Korea/S5/03(H9N2)) PB2 (PB2) gene, partial coding sequence.

gi|58429692|AY862713| Influenza A virus (A/Chicken/Korea/S12/03(H9N2)) PB2 (PB2) gene, partial coding sequence.

gi|58429694|AY862714| Influenza A virus (A/duck/Korea/S13/03(H9N2)) PB2 (PB2) gene, partial coding sequence.

gi|58429696|AY862715| Influenza A virus (A/pigeon/Korea/S14/03(H9N2)) PB2 (PB2) gene, partial coding sequence.

gi|58429698|AY862716| Influenza A virus (A/Chicken/Korea/S15/03(H9N2)) PB2 (PB2) gene, partial coding sequence.

gi|58429700|AY862717| Influenza A virus (A/Chicken/Korea/S16/03(H9N2)) PB2 (PB2) gene, partial coding sequence.

gi|58429702|AY862718| Influenza A virus (A/Chicken/Korea/S18/03(H9N2)) PB2 (PB2) gene, partial coding sequence.

Sequences Used in Influenza A Polymerase Acidic Protein (PA) Assays

gi|27465935|AY180662| Influenza A virus strain A/quail/Nanchang/12-340/2000 (H1N1) polymerase subunit PA (PA) gene, partial coding sequence.

gi|49357063|AY633217| Influenza A virus (A/Mallard/Alberta/211/98 (H1N1)) polymerase protein A (PA) gene, partial coding sequence.

gi|5918195|AJ243994| Partial mRNA of PA protein of influenza A virus (strain A/Mallard/New York/6750/78).

gi|45272157|AY422034| Influenza A virus (A/duck/Hokkaido/95/01(H2N2)) PA protein (PA) gene, partial coding sequence.

gi|27465965|AY180677| Influenza A virus strain A/chicken/Nanchang/3-120/2001 (H3N2) polymerase subunit PA (PA) gene, partial coding sequence.

gi|56159994|AY779263| Influenza A virus (A/Turkey/North Carolina/12344/03(H3N2)) polymerase acid protein (PA) gene, partial coding sequence.

gi|56159996|AY779264| Influenza A virus (A/Turkey/Minnesota/764-2/03(H3N2)) polymerase acid protein (PA) gene, partial coding sequence.

gi|58429736|AY862687| Influenza A virus (A/Chicken/Korea/S6/03(H3N2)) PA(PA) gene, partial coding sequence.

gi|58429738|AY862688| Influenza A virus (A/duck/Korea/S7/03(H3N2)) PA(PA) gene, partial coding sequence.

gi|58429740|AY862689| Influenza A virus (A/duck/Korea/S8/03(H3N2)) PA(PA) gene, partial coding sequence.

gi|58429742|AY862690| Influenza A virus (A/duck/Korea/S9/03(H3N2)) PA(PA) gene, partial coding sequence.

gi|58429744|AY862691| Influenza A virus (A/duck/Korea/S10/03(H3N2)) PA(PA) gene, partial coding sequence.

gi|58429746|AY862692| Influenza A virus (A/pigeon/Korea/S11/03(H3N2)) PA(PA) gene, partial coding sequence.

gi|18091833|AF213914| Influenza A virus (A/chicken/Italy/5945/95 (H3N2)) fragment 3PA polymerase protein gene, partial coding sequence.

gi|58531086|AB166861| Influenza A virus (A/Chicken/Yangkou/7/2004(H5N1)) polymerase acid protein PA gene, complete coding sequence.

gi|58531118|AB188815| Influenza A virus (A/chicken/Oita/8/2004(H5N1)) polymerase acid protein PA gene, complete coding sequence.

gi|9863935|AF216739| Influenza A virus (A/environment/Hong Kong/437-10/99(H5N1)) polymerase acid protein gene, complete coding sequence.

gi|14165201|AF380163| Influenza A virus (A/goose/Guangdong/3/97(H5N1)) fragment 3 polymerase (PA) gene, complete coding sequence.

gi|18092185|AF398427| Influenza A virus (A/goose/Hong Kong/385.3/2000 (H5N1)) polymerase (PA) gene, partial coding sequence.

gi|18092187|AF398428| Influenza A virus (A/goose/Hong Kong/385.5/2000 (H5N1)) polymerase (PA) gene, partial coding sequence.

gi|21359667|AF468841| Influenza A virus (A/duck/Anyang/AVL-1/2001(H5N1)) polymerase acid protein (PA) gene, partial coding sequence.

gi|28849710|AF509195| Influenza A virus (A/chicken/Hong Kong/FY77/01(H5N1)) polymerase (PA) gene, partial coding sequence.

gi|28849712|AF509196| Influenza A virus (A/chicken/Hong Kong/YU562/01(H5N1)) polymerase (PA) gene, partial coding sequence.

gi|28849714|AF509197| Influenza A virus (A/chick/Hong Kong/YU563/01(H5N1)) polymerase (PA) gene, complete coding sequence.

gi|28849716|AF509198| Influenza A virus (A/chicken/Hong Kong/FY150/01(H5N1)) polymerase (PA) gene, partial coding sequence.

gi|28849718|AF509199| Influenza A virus (A/pheasant/Hong Kong/FY155/01(H5N1)) polymerase (PA) gene, partial coding sequence.

gi|28849720|AF509200| Influenza A virus (A/Silkie/Hong Kong/SF189/01(H5N1)) polymerase (PA) gene, partial coding sequence.

gi|28849722|AF509201| Influenza A virus (A/quail/Hong Kong/SF203/01(H5N1)) polymerase (PA) gene, partial coding sequence.

gi|28849724|AF509202| Influenza A virus (A/pigeon/Hong Kong/SF215/01(H5N1)) polymerase(PA) gene, partial coding sequence.

gi|28849726|AF509203| Influenza A virus (A/chicken/Hong Kong/SF219/01(H5N1)) polymerase (PA) gene, partial coding sequence.

gi|28849728|AF509204| Influenza A virus (A/chicken/Hong Kong/715.5/01(H5N1)) polymerase (PA) gene, partial coding sequence.

gi|28849730|AF509205| Influenza A virus (A/chicken/Hong Kong/751.1/01(H5N1)) polymerase (PA) gene, partial coding sequence.

gi|28849732|AF509206| Influenza A virus (A/chicken/Hong Kong/822.1/01(H5N1)) polymerase (PA) gene, partial coding sequence.

gi|28849734|AF509207| Influenza A virus (A/chicken/Hong Kong/829.2/01(H5N1)) polymerase (PA) gene, partial coding sequence.

gi|28849736|AF509208| Influenza A virus (A/chicken/Hong Kong/830.2/01(H5N1)) polymerase (PA) gene, partial coding sequence.

gi|28849738|AF509209| Influenza A virus (A/chicken/Hong Kong/858.3/01(H5N1)) polymerase (PA) gene, partial coding sequence.

gi|28849740|AF509210| Influenza A virus (A/chicken/Hong Kong/866.3/01(H5N1)) polymerase (PA) gene, partial coding sequence.

gi|28849742|AF509211| Influenza A virus (A/chicken/Hong Kong/867.1/01(H5N1)) polymerase (PA) gene, partial coding sequence.

gi|28849744|AF509212| Influenza A virus (A/chicken/Hong Kong/879.1/01(H5N1)) polymerase (PA) gene, partial coding sequence.

gi|28849746|AF509213| Influenza A virus (A/chicken/Hong Kong/873.3/01(H5N1)) polymerase (PA) gene, partial coding sequence.

gi|28849748|AF509214| Influenza A virus (A/chicken/Hong Kong/876.1/01(H5N1)) polymerase (PA) gene, partial coding sequence.

gi|28849750|AF509215| Influenza A virus (A/chicken/Hong Kong/891.1/01(H5N1)) polymerase (PA) gene, partial coding sequence.

gi|28849752|AF509216| Influenza A virus (A/chicken/Hong Kong/893.1/01(H5N1)) polymerase (PA) gene, partial coding sequence.

gi|28849754|AF509217| Influenza A virus (A/goose/Hong Kong/76.1/01(H5N1)) polymerase (PA) gene, partial coding sequence.

gi|28849756|AF509218| Influenza A virus (A/goose/Hong Kong/ww100/01(H5N1)) polymerase (PA) gene, partial coding sequence.

gi|28849758|AF509219| Influenza A virus (A/duck/Hong Kong/573.4/01(H5N1)) polymerase (PA) gene, partial coding sequence.

gi|28849760|AF509220| Influenza A virus (A/duck/Hong Kong/646.3/01(H5N1)) polymerase (PA) gene, partial coding sequence.

gi|19697861|AY059526| Influenza A virus (A/goose/Hong Kong/ww26/2000(H5N1)) fragment 3 polymerase (PA) gene, partial coding sequence.

gi|19697863|AY059527| Influenza A virus (A/goose/Hong Kong/ww28/2000(H5N1)) fragment 3 polymerase (PA) gene, partial coding sequence.

gi|19697865|AY059528| Influenza A virus (A/duck/Hong Kong/ww381/2000(H5N1)) segment 3 polymerase (PA) gene, partial coding sequence.

gi|19697869|AY059530| Influenza A virus (A/duck/Hong Kong/ww461/2000(H5N1)) fragment 3 polymerase (PA) gene, partial coding sequence.

gi|19697871|AY059531| Influenza A virus (A/goose/Hong Kong/ww491/2000(H5N1)) fragment 3 polymerase (PA) gene, partial coding sequence.

gi|19697873|AY059532| Influenza A virus (A/duck/Hong Kong/2986.1/2000(H5N1)) fragment 3 polymerase (PA) gene, partial coding sequence.

gi|19697875|AY059533| Influenza A virus (A/goose/Hong Kong/3014.8/2000(H5N1)) fragment 3 polymerase (PA) gene, partial coding sequence.

gi|28821204|AY221566| Influenza A virus (A/chicken/Hong Kong/NT873.3/01-MB(H5N1)) polymerase (PA) gene, partial coding sequence.

gi|28821206|AY221567| Influenza A virus (A/chicken/Hong Kong/NT873.3/01(H5N1)) polymerase (PA) gene, partial coding sequence.

gi|28821208|AY221568| Influenza A virus (A/chicken/Hong Kong/FY150/01-MB(H5N1)) polymerase (PA) gene, partial coding sequence.

gi|28821210|AY221569| Influenza A virus (A/Chick/Hong Kong/FY150/01(H5N1)) polymerase (PA) gene, partial coding sequence.

gi|28821212|AY221570| Influenza A virus (A/pheasant/Hong Kong/FY155/01-MB(H5N1)) polymerase (PA) gene, partial coding sequence.

gi|28821214|AY221571| Influenza A virus (A/pheasant/Hong Kong/FY155/01(H5N1)) polymerase (PA) gene, partial coding sequence.

gi|28821216|AY221572| Influenza A virus (A/chicken/Hong Kong/YU822.2/01-MB(H5N1)) polymerase (PA) gene, partial coding sequence.

gi|28821218|AY221573| Influenza A virus (A/chicken/Hong Kong/YU822.2/01(H5N1)) polymerase (PA) gene, partial coding sequence.

gi|28821220|AY221574| Influenza A virus (A/chicken/Hong Kong/YU562/01(H5N1)) polymerase (PA) gene, partial coding sequence.

gi|41207483|AY518365| Influenza A virus (A/Duck/China/E319-2/03(H5N1)) polymerase (PA) gene, complete coding sequence.

gi|51094114|AY551934| Influenza A virus (A/chick/Nakhon Pathom/Thailand/CU-K2/04(H5N1)) polymerase (PA) gene, complete coding sequence

gi|47834839|AY576406| Influenza A virus (A/Gs/Hong Kong/739.2/02(H5N1)) polymerase (PA) gene, partial coding sequence.

gi|47834841|AY576407| Influenza A virus (A/Eg/Hong Kong/757.3/02(H5N1)) polymerase (PA) gene, partial coding sequence.

gi|47834843|AY576408| Influenza A virus (A/G.H/Hong Kong/793.1/02(H5N1)) polymerase (PA) gene, partial coding sequence.

gi|47834845|AY576409| Influenza A virus (A/Dk/Hong Kong/821/02(H5N1)) polymerase (PA) gene, partial coding sequence.

gi|47834847|AY576410| Influenza A virus (A/Ck/Hong Kong/31.4/02(H5N1)) polymerase (PA) gene, complete coding sequence.

gi|47834849|AY576411| Influenza A virus (A/Ck/Hong Kong/61.9/02(H5N1)) polymerase (PA) gene, complete coding sequence.

gi|47834851|AY576412| Influenza A virus (A/Ck/Hong Kong/YU777/02(H5N1)) polymerase (PA) gene, complete coding sequence.

gi|47834853|AY576413| Influenza A virus (A/Ck/Hong Kong/96.1/02(H5N1)) polymerase (PA) gene, partial coding sequence.

gi|47834855|AY576414| Influenza A virus (A/Ck/Hong Kong/409.1/02(H5N1)) polymerase (PA) gene, partial coding sequence.

gi|47834857|AY576415| Influenza A virus (A/Ph/Hong Kong/sv674.15/02(H5N1)) polymerase (PA) gene, complete coding sequence.

gi|47156478|AY585462| Influenza A virus (A/duck/Fujian/01/2002(H5N1)) polymerase (PA) mRNA, complete coding sequence.

gi|47156480|AY585463| Influenza A virus (A/duck/Fujian/13/2002(H5N1)) polymerase (PA) mRNA, complete coding sequence.

gi|47156482|AY585464| Influenza A virus (A/duck/Fujian/17/2001(H5N1)) polymerase (PA) mRNA, complete coding sequence.

gi|47156484|AY585465| Influenza A virus (A/duck/Fujian/19/2000(H5N1)) polymerase (PA) mRNA, complete coding sequence.

gi|47156486|AY585466| Influenza A virus (A/duck/Guangdong/01/2001(H5N1)) polymerase (PA) mRNA, complete coding sequence.

gi|47156488|AY585467| Influenza A virus (A/duck/Guangdong/07/2000(H5N1)) polymerase (PA) mRNA, complete coding sequence.

gi|47156490|AY585468| Influenza A virus (A/duck/Guangdong/12/2000(H5N1)) polymerase (PA) mRNA, complete coding sequence.

gi|47156492|AY585469| Influenza A virus (A/duck/Guangdong/22/2002 (H5N1)) polymerase (PA) mRNA, complete coding sequence.

gi|47156494|AY585470| Influenza A virus (A/duck/Guangdong/40/2000(H5N1)) polymerase (PA) mRNA, complete coding sequence.

gi|47156496|AY585471| Influenza A virus (A/duck/Guangxi/07/1999 (H5N1)) polymerase (PA) mRNA, complete coding sequence.

gi|47156498|AY585472| Influenza A virus (A/duck/Guangxi/22/2001(H5N1)) polymerase (PA) mRNA, complete coding sequence.

gi|47156500|AY585473| Influenza A virus (A/duck/Guangxi/35/2001 (H5N1)) polymerase (PA) mRNA, complete coding sequence.

gi|47156502|AY585474| Influenza A virus (A/duck/Guangxi/50/2001(H5N1)) polymerase (PA) mRNA, complete coding sequence.

gi|47156504|AY585475| Influenza A virus (A/duck/Guangxi/53/2002(H5N1)) polymerase (PA) mRNA, complete coding sequence.

gi|47156506|AY585476| Influenza A virus (A/duck/Shanghai/08/2001(H5N1)) polymerase (PA) mRNA, complete coding sequence.

gi|47156508|AY585477| Influenza A virus (A/duck/Shanghai/13/2001(H5N1)) polymerase (PA) mRNA, complete coding sequence.

gi|47156510|AY585478| Influenza A virus (A/duck/Shanghai/35/2002 (H5N1)) polymerase (PA) mRNA, complete coding sequence.

gi|47156512|AY585479| Influenza A virus (A/duck/Shanghai/37/2002(H5N1)) polymerase (PA) mRNA, complete coding sequence.

gi|47156514|AY585480| Influenza A virus (A/duck/Shanghai/38/2001(H5N1)) polymerase (PA) mRNA, complete coding sequence.

gi|47156516|AY585481| Influenza A virus (A/duck/Zhejiang/11/2000 (H5N1)) polymerase (PA) mRNA, complete coding sequence.

gi|47156518|AY585482| Influenza A virus (A/duck/Zhejiang/52/2000(H5N1)) polymerase (PA) mRNA, complete coding sequence.

gi|47716770|AY609311| Influenza A virus (A/chicken/Guangdong/174/04(H5N1)) fragment 3, complete sequence.

gi|50313026|AY651597| Influenza A virus (A/Ck/Indonesia/4/2004 (H5N1)) polymerase acid protein (PA) gene, partial coding sequence.

gi|50313028|AY651598| Influenza A virus (A/Ck/Indonesia/5/2004(H5N1)) polymerase acid protein (PA) gene, partial coding sequence.

gi|50313030|AY651599| Influenza A virus (A/Ck/Indonesia/2A/2003(H5N1)) polymerase acid protein (PA) gene, partial coding sequence.

gi|50313032|AY651600| Influenza A virus (A/Dk/Indonesia/MS/2004(H5N1)) polymerase acid protein (PA) gene, partial coding sequence.

gi|50313034|AY651601| Influenza A virus (A/Ck/Indonesia/BL/2003(H5N1)) polymerase acid protein (PA) gene, partial coding sequence.

gi|50313036|AY651602| Influenza A virus (A/Ck/Indonesia/PA/2003(H5N1)) polymerase acid protein (PA) gene, partial coding sequence.

gi|50313038|AY651603| Influenza A virus (A/Ck/Thailand/1/2004(H5N1)) polymerase acid protein (PA) gene, partial coding sequence.

gi|50313040|AY651604| Influenza A virus (A/Ck/Thailand/73/2004(H5N1)) polymerase acid protein (PA) gene, partial coding sequence.

gi|50313042|AY651605| Influenza A virus (A/Ck/Thailand/9.1/2004(H5N1)) polymerase acid protein (PA) gene, partial coding sequence.

gi|50313044|AY651606| Influenza A virus (A/Qa/Thailand/57/2004(H5N1)) polymerase acid protein (PA) gene, partial coding sequence.

gi|50313046|AY651607| Influenza A virus (A/bird/Thailand/3.1/2004(H5N1)) polymerase acid protein (PA) gene, partial coding sequence.

gi|50313048|AY651608| Influenza A virus (A/Dk/Thailand/71.1/2004 (H5N1)) polymerase acid protein (PA) gene, partial coding sequence.

gi|50313050|AY651609| Influenza A virus (A/Gs/Thailand/79/2004(H5N1)) polymerase acid protein (PA) gene, partial coding sequence.

gi|50313060|AY651614| Influenza A virus (A/Ck/Vietnam/33/2004(H5N1)) polymerase acid protein (PA) gene, partial coding sequence.

gi|50313062|AY651615| Influenza A virus (A/Ck/Vietnam/35/2004(H5N1)) polymerase acid protein (PA) gene, partial coding sequence.

gi|50313064|AY651616| Influenza A virus (A/Ck/Vietnam/36/2004(H5N1)) polymerase acid protein (PA) gene, partial coding sequence.

gi|50313066|AY651617| Influenza A virus (A/Ck/Vietnam/37/2004 (H5N1)) polymerase acid protein (PA) gene, partial coding sequence.

gi|50313068|AY651618| Influenza A virus (A/Ck/Vietnam/38/2004(H5N1)) polymerase acid protein (PA) gene, partial coding sequence.

gi|50313070|AY651619| Influenza A virus (A/Ck/Vietnam/39/2004(H5N1)) polymerase acid protein (PA) gene, partial coding sequence.

gi|50313072|AY651620| Influenza A virus (A/Ck/Vietnam/C57/2004(H5N1)) polymerase acid protein (PA) gene, partial coding sequence.

gi|50313074|AY651621| Influenza A virus (A/Dk/Vietnam/11/2004(H5N1)) polymerase acid protein (PA) gene, partial coding sequence.

gi|50313076|AY651622| Influenza A virus (A/Gf/Hong Kong/38/2002 (H5N1)) polymerase acid protein (PA) gene, complete coding sequence.

gi|50313078|AY651623| Influenza A virus (A/Ck/Hong Kong/31.2/2002 (H5N1)) polymerase acid protein (PA) gene, complete coding sequence.

gi|50313080|AY651624| Influenza A virus (A/Ck/Hong Kong/37.4/2002 (H5N1)) polymerase acid protein (PA) gene, partial coding sequence.

gi|50313082|AY651625| Influenza A virus (A/SCk/Hong Kong/YU100/2002(H5N1)) polymerase acid protein (PA) gene, complete coding sequence.

gi|50313084|AY651626| Influenza A virus (A/Ck/Hong Kong/YU22/2002(H5N1)) polymerase acid protein (PA) gene, partial coding sequence.

gi|50313086|AY651627| Influenza A virus (A/Ck/Hong Kong/3176.3/2002 (H5N1)) polymerase acid protein (PA) gene, partial coding sequence.

gi|50313088|AY651628| Influenza A virus (A/Ck/Hong Kong/3169.1/2002 (H5N1)) polymerase acid protein (PA) gene, partial coding sequence.

gi|50313090|AY651629| Influenza A virus (A/Ck/Hong Kong/FY157/2003(H5N1)) polymerase acid protein (PA) gene, partial coding sequence.

gi|50313092|AY651630| Influenza A virus (A/Ck/Hong Kong/YU324/2003(H5N1)) polymerase acid protein (PA) gene, partial coding sequence.

gi|50313094|AY651631| Influenza A virus (A/Ck/Hong Kong/2133.1/2003 (H5N1)) polymerase acid protein (PA) gene, partial coding sequence.

gi|50313096|AY651632| Influenza A virus (A/Ck/Hong Kong/NT93/2003 (H5N1)) polymerase acid protein (PA) gene, partial coding sequence.

gi|50313098|AY651633| Influenza A virus (A/Ck/Hong Kong/SSP141/2003(H5N1)) polymerase acid protein (PA) gene, partial coding sequence.

gi|50313100|AY651634| Influenza A virus (A/Ck/Hong Kong/WF157/2003(H5N1)) polymerase acid protein (PA) gene, partial coding sequence.

gi|50313102|AY651635| Influenza A virus (A/Red-headed Gull/Hong Kong/12.1/2003 (H5N1)) polymerase acid protein (PA) gene, partial coding sequence.

gi|50313104|AY651636| Influenza A virus (A/heron/Hong Kong/861.1/2002 (H5N1)) polymerase acid protein (PA) gene, partial coding sequence.

gi|50313106|AY651637| Influenza A virus (A/Dove/Hong Kong/862.7/2002 (H5N1)) polymerase acid protein (PA) gene, partial coding sequence.

gi|50313108|AY651638| Influenza A virus (A/Tree Finch/Hong Kong/864/2002 (H5N1)) polymerase acid protein (PA) gene, partial coding sequence.

gi|50313110|AY651639| Influenza A virus (A/teal/China/2978.1/2002(H5N1)) polymerase acid protein (PA) gene, partial coding sequence.

gi|50313112|AY651640| Influenza A virus (A/Peregrine/Hong Kong/D0028/2004(H5N1)) polymerase acid protein (PA) gene, partial coding sequence.

gi|50313114|AY651641| Influenza A virus (A/Dk/HN/5806/2003 (H5N1)) polymerase acid protein (PA) gene, partial coding sequence.

gi|50313116|AY651642| Influenza A virus (A/Dk/HN/303/2004 (H5N1)) polymerase acid protein (PA) gene, partial coding sequence.

gi|50313118|AY651643| Influenza A virus (A/Dk/HN/101/2004 (H5N1)) polymerase acid protein (PA) gene, partial coding sequence.

gi|50313120|AY651644| Influenza A virus (A/Dk/ST/4003/2003 (H5N1)) polymerase acid protein (PA) gene, partial coding sequence.

gi|50313122|AY651645| Influenza A virus (A/Ph/ST/44/2004 (H5N1)) polymerase acid protein (PA) gene, partial coding sequence.

gi|50313124|AY651646| Influenza A virus (A/Ck/ST/4231/2003 (H5N1)) polymerase acid protein (PA) gene, partial coding sequence.

gi|50313126|AY651647| Influenza A virus (A/Dk/YN/6255/2003 (H5N1)) polymerase acid protein (PA) gene, partial coding sequence.

gi|50313128|AY651648| Influenza A virus (A/Dk/YN/6445/2003(H5N1)) polymerase acid protein (PA) gene, partial coding sequence.

gi|50313130|AY651649| Influenza A virus (A/Ck/YN/115/2004 (H5N1)) polymerase acid protein (PA) gene, partial coding sequence.

gi|50313132|AY651650| Influenza A virus (A/Ck/YN/374/2004 (H5N1)) polymerase acid protein (PA) gene, partial coding sequence.

gi|50365724|AY653198| Influenza A virus (A/chicken/Jilin/9/2004(H5N1)) segment 3, complete sequence.

gi|56548923|AY676029| Influenza A virus (A/duck/Hong Kong/821/02(H5N1)) polymerase (PA) gene, complete coding sequence.

gi|56548925|AY676030| Influenza A virus (A/Egret/Hong Kong/757.2/03(H5N1)) polymerase (PA) gene, complete coding sequence.

gi|56548927|AY676031| Influenza A virus (A/chick/Korea/ES/03(H5N1)) polymerase (PA) gene, complete coding sequence.

gi|56548929|AY676032| Influenza A virus (A/Duck/Korea/ESDI/03(H5N1)) polymerase (PA) gene, complete coding sequence.

gi|50956625|AY684705| Influenza A virus (A/chicken/Hubei/327/2004 (H5N1)) polymerase (PA) gene, complete coding sequence.

gi|56119221|AY720944| Influenza A virus (A/chicken/Vietnam/DT-171/2004(H5N1)) polymerase acid protein (PA) gene, partial coding sequence.

gi|56119227|AY720947| Influenza A virus (A/duck/Vietnam/TG-007A/2004(H5N1)) polymerase acid protein (PA) gene, partial coding sequence.

gi|57924419|AY724784| Influenza A virus (A/chicken/Vietnam/HCM-022/2004(H5N1)) polymerase (PA) gene, partial coding sequence.

gi|57924480|AY724786| Influenza A virus (A/chicken/Vietnam/DN-045/2004(H5N1)) polymerase (PA) gene, partial coding sequence.

gi|57924569|AY724788| Influenza A virus (A/chicken/Vietnam/VL-008/2004(H5N1)) polymerase (PA) gene, partial coding sequence.

gi|57924680|AY724790| Influenza A virus (A/chicken/Vietnam/AG-010/2004(H5N1)) polymerase (PA) gene, partial coding sequence.

gi|57924765|AY724792| Influenza A virus (A/chicken/Vietnam/DT-015/2004(H5N1)) polymerase (PA) gene, partial coding sequence.

gi|57924882|AY724796| Influenza A virus (A/chicken/Vietnam/LA-024/2004(H5N1)) polymerase (PA) gene, partial coding sequence.

gi|57915971|AY737288| Influenza A virus (A/chicken/Guangdong/191/04(H5N1)) segment 3, complete sequence.

gi|57916018|AY737295| Influenza A virus (A/chicken/Guangdong/178/04(H5N1)) segment 3, complete sequence.

gi|57916074|AY737303| Influenza A virus (A/duck/Guangdong/173/04(H5N1)) segment 3, complete sequence.

gi|55233234|AY770082| Influenza A virus (A/chicken/Hubei/489/2004 (H5N1)) polymerase (PA) gene, complete coding sequence.

gi|54873465|AY770995| Influenza A virus (A/chick/Ayutthaya/Thailand/CU-23/04(H5N1)) polymerase gene, partial coding sequence.

gi|58618433|AY818133| Influenza A virus (A/chicken/Vietnam/C58/04(H5N1)) polymerase protein PA gene, complete coding sequence.

gi|58618435|AY818134| Influenza A virus (A/quail/Vietnam/36/04(H5N1)) polymerase protein PA gene, complete coding sequence.

gi|58374187|AY856863| Influenza A virus (A/duck/Shandong/093/2004(H5N1)) segment 3, complete sequence.

gi|58531136|AB188823| Influenza A virus (A/chicken/Kyoto/3/2004(H5N1)) polymerase acid protein PA gene, complete coding sequence.

gi|58531154|AB189052| Influenza A virus (A/crow/Kyoto/53/2004(H5N1)) polymerase acid protein PA gene, complete coding sequence, .

gi|58531170|AB189059| Influenza A virus (A/crow/Osaka/102/2004 (H5N1)) polymerase acid protein PA gene, complete coding sequence.

gi|3335418|AF046087| Influenza A virus (A/chicken/Hong Kong/220/97(H5N1)) polymerase acid protein (PA) gene, partial coding sequence.

gi|6048895|AF098604| Influenza A virus (A/Chick/Hong Kong/258/97(H5N1)) PA protein (PA) gene, complete coding sequence.

gi|6048897|AF098605| Influenza A virus (A/Chick/Hong Kong/y388/97(H5N1)) PA protein (PA) gene, complete coding sequence.

gi|6048899|AF098606| Influenza A virus (A/Chicken/Hong Kong/728/97(H5N1)) PA protein (PA) gene, complete coding sequence.

gi|6048901|AF098607| Influenza A virus (A/Chicken/Hong Kong/786/97(H5N1)) PA protein (PA) gene, complete coding sequence.

gi|6048903|AF098608| Influenza A virus (A/Chicken/Hong Kong/915/97(H5N1)) PA protein (PA) gene, complete coding sequence.

gi|6048905|AF098609| Influenza A virus (A/duck/Hong Kong/p46/97(H5N1)) PA protein (PA) gene, complete coding sequence.

gi|6048907|AF098610| Influenza A virus (A/duck/Hong Kong/y283/97(H5N1)) PA protein (PA) gene, complete coding sequence.

gi|6048909|AF098611| Influenza A virus (A/goose/Hong Kong/w355/97(H5N1)) PA protein (PA) gene, complete coding sequence.

gi|5805280|AF144302| Influenza A virus (A/goose/Guangdong/1/96(H5N1)) polymerase (PA) gene, complete coding sequence.

gi|9863880|AF216715| Influenza A virus (A/environment/Hong Kong/437-4/99(H5N1)) polymerase acid protein gene, complete coding sequence.

gi|9863899|AF216723| Influenza A virus (A/environment/Hong Kong/437-6/99(H5N1)) polymerase acid protein gene, complete coding sequence.

gi|9863917|AF216731| Influenza A virus (A/environment/Hong Kong/437-8/99(H5N1)) polymerase acid protein gene, complete coding sequence.

gi|34597776|AY303660| Influenza A virus (A/Chicken/Chile/176822/02(H7N3)) polymerase acid protein gene, complete coding sequence.

gi|34597778|AY303661| Influenza A virus (A/Chicken/Chile/4957/02(H7N3)) polymerase acid protein gene, complete coding sequence.

gi|34597780|AY303662| Influenza A virus (A/Chicken/Chile/4322/02(H7N3)) polymerase acid protein gene, partial coding sequence.

gi|47680912|AY586431| Influenza A virus (A/Duck/Italy/43/01(H7N3)) PA gene, partial coding sequence.

gi|47680914|AY586432| Influenza A virus (A/Duck/Italy/33/01(H7N3)) PA gene, partial coding sequence.

gi|47680916|AY586433| Influenza A virus (A/Turkey/Italy/220158/2002(H7N3)) PA gene, partial coding sequence.

gi|47680918|AY586434| Influenza A virus (A/Turkey/Italy/214845/02(H7N3)) PA gene, partial coding sequence.

gi|47834370|AY616764| Influenza A virus (A/chick/BC/04(H7N3)) polymerase acid protein 2 (PA) gene, complete coding sequence.

gi|50542647|AY646083| Influenza A virus (A/Chicken/British Columbia/GSC_Human_B/04(H7N3)) polymerase acid protein 2 (PA) gene, complete coding sequence.

gi|50083049|AY648292| Influenza A virus (A/GSC_chick_B/British Columbia/04(H7N3)) polymerase acid protein 2 (PA) gene, complete coding sequence.

gi|50059192|AY650275| Influenza A virus (A/GSC_Chicken/British Columbia/04(H7N3)) polymerase acid protein 2 (PA) gene, complete coding sequence.

gi|9988639|AF268109| Influenza A virus (A/Retknot/Delaware/259/94(H7N7)) polymerase protein PA gene, partial coding sequence.

gi|40353080|AJ619677| Influenza A virus (A/chicken/Germany/R28/03(H7N7)) polymerase complex subunit (PA) PA gene, genomic RNA.

gi|37813167|AY342415| Influenza A virus (A/Netherlands/124/03(H7N7)) polymerase protein A gene, partial coding sequence.

gi|37813169|AY342416| Influenza A virus (A/Netherlands/126/03(H7N7)) polymerase protein A gene, partial coding sequence.

gi|37813171|AY342417| Influenza A virus (A/Netherlands/127/03(H7N7)) polymerase protein A gene, partial coding sequence.

gi|37813173|AY342418| Influenza A virus (A/Netherlands/219/03(H7N7)) polymerase protein A gene, complete coding sequence.

gi|37813175|AY342419| Influenza A virus (A/Netherlands/033/03(H7N7)) polymerase protein A gene, complete coding sequence.

gi|37813177|AY342420| Influenza A virus (A/chick/Netherlands/1/03(H7N7)) polymerase protein A gene, complete coding sequence.

gi|13383274|AB049157| Influenza A virus (A/parakeet/Chiba/1/97(H9N2)) polymerase acid protein PA gene, complete coding sequence.

gi|13383276|AB049158| Influenza A virus (A/parakeet/Narita/92A/98(H9N2)) polymerase acid protein PA gene, complete coding sequence.

gi|33318154|AF508662| Influenza A virus (A/ostrich/South Africa/9508103/95 (H9N2)) segment 3 polymerase PA (PA) gene, complete coding sequence.

gi|33318156|AF508663| Influenza A virus (A/chick/Pakistan/4/99(H9N2)) segment 3 polymerase PA (PA) gene, complete coding sequence.

gi|33318158|AF508664| Influenza A virus (A/chicken/Pakistan/5/99(H9N2)) segment 3 polymerase PA (PA) gene, partial coding sequence.

gi|33318160|AF508665| Influenza A virus (A/chicken/Germany/R45/98(H9N2)) segment 3 polymerase PA (PA) gene, complete coding sequence.

gi|33318162|AF508666| Influenza A virus (A/Duck/Germany/113/95(H9N2)) fragment 3 polymerase PA (PA) gene, complete coding sequence.

gi|33318164|AF508667| Influenza A virus (A/chicken/Iran/11T/99(H9N2)) segment 3 polymerase PA (PA) gene, partial coding sequence.

gi|33318166|AF508668| Influenza A virus (A/Chicken/Saudi Arabia/532/99(H9N2)) segment 3 polymerase PA (PA) gene, complete coding sequence.

gi|33318168|AF508669| Influenza A virus (A/pheasant/Ireland/PV18/97(H9N2)) fragment 3 polymerase PA (PA) gene, complete coding sequence.

gi|33318170|AF508670| Influenza A virus (A/Chicken/Korea/99029/99(H9N2)) segment 3 polymerase PA (PA) gene, complete coding sequence.

gi|33318172|AF508671| Influenza A virus (A/chicken/Beijing/8/98(H9N2)) fragment 3 polymerase PA (PA) gene, complete coding sequence.

gi|33318174|AF508672| Influenza A virus (A/chicken/Guangdong/10/00(H9N2)) fragment 3 polymerase PA (PA) gene, complete coding sequence.

gi|33318176|AF508673| Influenza A virus (A/Chicken/Guangdong/11/97(H9N2)) segment 3 polymerase PA (PA) gene, complete coding sequence.

gi|33318178|AF508674| Influenza A virus (A/chicken/Hebei/4/98(H9N2)) fragment 3 polymerase PA (PA) gene, complete coding sequence.

gi|33318180|AF508675| Influenza A virus (A/Chicken/Heilongjiang/10/97(H9N2)) fragment 3 polymerase PA (PA) gene, complete coding sequence.

gi|33318182|AF508676| Influenza A virus (A/chicken/Henan/62/00(H9N2)) segment 3 polymerase PA (PA) gene, partial coding sequence.

gi|33318184|AF508677| Influenza A virus (A/chicken/Ningxia/5/99 (H9N2)) fragment 3 polymerase PA (PA) gene, complete coding sequence.

gi|33318186|AF508678| Influenza A virus (A/Chicken/Sichuan/5/97(H9N2)) segment 3 polymerase PA (PA) gene, complete coding sequence.

gi|33318188|AF508679| Influenza A virus (A/chicken/Shandong/6/96(H9N2)) fragment 3 polymerase PA (PA) gene, partial coding sequence.

gi|33318190|AF508680| Influenza A virus (A/chicken/Shijiazhuang/2/99 (H9N2)) segment 3 polymerase PA (PA) gene, partial coding sequence.

gi|33318192|AF508681| Influenza A virus (A/chicken/Shenzhen/9/97(H9N2)) segment 3 polymerase PA (PA) gene, complete coding sequence.

gi|33318194|AF508682| Influenza A virus (A/duck/Nanjing/1/97 (H9N2)) segment 3 polymerase PA (PA) gene, complete coding sequence.

gi|33318196|AF508683| Influenza A virus (A/quail/Shanghai/8/96 (H9N2)) fragment 3 polymerase PA (PA) gene, partial coding sequence.

gi|31339541|AF523446| Influenza A virus (A/duck/Shantou/1043/00(H9N2)) polymerase (PA) gene, partial coding sequence.

gi|31339543|AF523447| Influenza A virus (A/duck/Shantou/1042/00(H9N2)) polymerase (PA) gene, partial coding sequence.

gi|31339545|AF523448| Influenza A virus (A/duck/Shantou/2088/01(H9N2)) polymerase (PA) gene, partial coding sequence.

gi|31339547|AF523449| Influenza A virus (A/duck/Shantou/830/00(H9N2)) polymerase (PA) gene, partial coding sequence.

gi|31339549|AF523450| Influenza A virus (A/duck/Shantou/1796/00(H9N2)) polymerase (PA) gene, partial coding sequence.

gi|31339551|AF523451| Influenza A virus (A/duck/Shantou/2143/00(H9N2)) polymerase (PA) gene, partial coding sequence.

gi|31339553|AF523452| Influenza A virus (A/duck/Shantou/2134/00(H9N2)) polymerase (PA) gene, partial coding sequence.

gi|31339555|AF523453| Influenza A virus (A/duck/Shantou/2144/00(H9N2)) polymerase (PA) gene, partial coding sequence.

gi|31339557|AF523454| Influenza A virus (A/wild duck/Shantou/4808/01(H9N2)) polymerase (PA) gene, partial coding sequence.

gi|31339559|AF523455| Influenza A virus (A/duck/Shantou/1881/00(H9N2)) polymerase (PA) gene, partial coding sequence.

gi|31339561|AF523456| Influenza A virus (A/duck/Shantou/2102/00(H9N2)) polymerase (PA) gene, partial coding sequence.

gi|31339563|AF523457| Influenza A virus (A/duck/Hong Kong/289/78 (H9N2)) polymerase (PA) gene, partial coding sequence.

gi|31339565|AF523458| Influenza A virus (A/duck/Hong Kong/610/79(H9N2)) polymerase (PA) gene, partial coding sequence.

gi|31339567|AF523459| Influenza A virus (A/duck/Hong Kong/86/76(H9N2)) polymerase (PA) gene, partial coding sequence.

gi|31339569|AF523460| Influenza A virus (A/duck/Hong Kong/366/78 (H9N2)) polymerase (PA) gene, partial coding sequence.

gi|22759040|AF536669| Influenza A virus (A/chicken/Beijing/1/95(H9N2)) PA gene, partial coding sequence.

gi|22759042|AF536670| Influenza A virus (A/chicken/Beijing/2/97(H9N2)) PA gene, partial coding sequence.

gi|22759044|AF536671| Influenza A virus (A/chicken/Beijing/3/99(H9N2)) PA gene, partial coding sequence.

gi|22759046|AF536672| Influenza A virus (A/chicken/Guangdong/97(H9N2)) PA gene, partial coding sequence.

gi|22759048|AF536673| Influenza A virus (A/chicken/Hebei/1/96(H9N2)) PA gene, partial coding sequence.

gi|22759050|AF536674| Influenza A virus (A/chicken/Hebei/2/98(H9N2)) PA gene, partial coding sequence.

gi|22759052|AF536675| Influenza A virus (A/chicken/Hebei/3/98(H9N2)) PA gene, partial coding sequence.

gi|22759054|AF536676| Influenza A virus (A/Chicken/Henan/98(H9N2)) PA gene, partial coding sequence.

gi|22759056|AF536677| Influenza A virus (A/chicken/Liaoning/99(H9N2)) PA gene, partial coding sequence.

gi|22759058|AF536678| Influenza A virus (A/Chicken/Shandong/98(H9N2)) PA gene, partial coding sequence.

gi|12038897|AJ291397| Influenza A virus (A/chick/Pakistan/2/99(H9N2)) polymerase (PA) PA gene, genomic RNA.

gi|18496121|AJ427863| Influenza A virus (A/quail/Hong Kong/FY298/00(H9N2)) PA polymerase protein part pa gene, genomic RNA

gi|27465911|AY180650| Influenza A virus strain A/duck/Nanchang/11-392/2000 (H9N2) polymerase subunit PA (PA) gene, partial coding sequence.

gi|27465913|AY180651| Influenza A virus strain A/duck/Nanchang/11-290/2000 (H9N2) polymerase subunit PA (PA) gene, partial coding sequence.

gi|27465915|AY180652| Influenza A virus strain A/chicken/Nanchang/1-0016/2000 (H9N2) polymerase subunit PA (PA) gene, partial coding sequence.

gi|27465917|AY180653| Influenza A virus strain A/duck/Nanchang/11-197/2000 (H9N2) polymerase subunit PA (PA) gene, partial coding sequence.

gi|27465937|AY180663| Influenza A virus strain A/pigeon/Nanchang/2-0461/2000 (H9N2) polymerase subunit PA (PA) gene, partial coding sequence.

gi|27465941|AY180665| Influenza A virus strain A/chicken/Nanchang/4-301/2001 (H9N2) polymerase subunit PA (PA) gene, partial coding sequence.

gi|27465943|AY180666| Influenza A virus strain A/chicken/Nanchang/4-361/2001 (H9N2) polymerase subunit PA (PA) gene, partial coding sequence.

gi|27465961|AY180675| Influenza A virus strain A/wild duck/Nanchang/2-0480/2000 (H9N2) polymerase subunit PA (PA) gene, partial coding sequence.

gi|27465983|AY180686| Influenza A virus strain A/duck/Nanchang/1-0070/2000 (H9N2) polymerase subunit PA (PA) gene, partial coding sequence.

gi|27465989|AY180689| Influenza A virus strain A/duck/Nanchang/10-389/2000 (H9N2) polymerase subunit PA (PA) gene, partial coding sequence.

gi|27465993|AY180691| Influenza A virus strain A/pigeon/Nanchang/11-145/2000 (H9N2) polymerase subunit PA (PA) gene, partial coding sequence.

gi|27466001|AY180695| Influenza A virus strain A/quail/Nanchang/2-0460/2000 (H9N2) polymerase subunit PA (PA) gene, partial coding sequence.

gi|27466003|AY180696| Influenza A virus strain A/quail/Nanchang/4-040/2000 (H9N2) polymerase subunit PA (PA) gene, partial coding sequence.

gi|27466009|AY180699| Influenza A virus strain A/chicken/Nanchang/4-010/2000 (H9N2) polymerase subunit PA (PA) gene, partial coding sequence.

gi|27466015|AY180702| Influenza A virus strain A/duck/Nanchang/7-092/2000 (H9N2) polymerase subunit PA (PA) gene, partial coding sequence.

gi|27466019|AY180704| Influenza A virus strain A/pigeon/Nanchang/7-058/2000 (H9N2) polymerase subunit PA (PA) gene, partial coding sequence.

gi|30025973|AY253752| Influenza A virus (A/chicken/Shanghai/F/98(H9N2)) polymerase acid protein (PA) gene, complete coding sequence.

gi|49357051|AY633169| Influenza A virus (A/Mallard/Alberta/17/91 (H9N2)) polymerase protein A (PA) gene, partial coding sequence.

gi|49357079|AY633281| Influenza A virus (A/Mallard/Alberta/321/88 (H9N2)) polymerase protein A (PA) gene, partial coding sequence.

gi|49357083|AY633297| Influenza A virus (A/Mallard/Alberta/11/91(H9N2)) polymerase protein A (PA) gene, partial coding sequence.

gi|54301528|AY664755| Influenza A virus (A/chicken/Hong Kong/CSW153/03(H9N2)) polymerase acid protein (PA) gene, partial coding sequence.

gi|54301530|AY664756| Influenza A virus (A/chicken/Hong Kong/AP45/03(H9N2)) polymerase acid protein (PA) gene, partial coding sequence.

gi|54301532|AY664757| Influenza A virus (A/chicken/Hong Kong/BD90/03(H9N2)) polymerase acid protein (PA) gene, partial coding sequence.

gi|54301534|AY664758| Influenza A virus (A/chicken/Hong Kong/CSW291/03(H9N2)) polymerase acid protein (PA) gene, partial coding sequence.

gi|54301536|AY664759| Influenza A virus (A/chicken/Hong Kong/CSW304/03(H9N2)) polymerase acid protein (PA) gene, partial coding sequence.

gi|54301538|AY664760| Influenza A virus (A/chicken/Hong Kong/FY23/03(H9N2)) polymerase acid protein (PA) gene, partial coding sequence.

gi|54301540|AY664761| Influenza A virus (A/Grizzly chicken/Hong Kong/NT10I/03(H9N2)) polymerase acidic protein (PA) gene, partial coding sequence.

gi|54301542|AY664762| Influenza A virus (A/chicken/Hong Kong/NT142/03(H9N2)) polymerase acid protein (PA) gene, partial coding sequence.

gi|54301544|AY664763| Influenza A virus (A/chicken/Hong Kong/SF1/03(H9N2)) polymerase acid protein (PA) gene, partial coding sequence.

gi|54301546|AY664764| Influenza A virus (A/chicken/Hong Kong/SSP101/03(H9N2)) polymerase acid protein (PA) gene, partial coding sequence.

gi|54301548|AY664765| Influenza A virus (A/chicken/Hong Kong/TP38/03(H9N2)) polymerase acid protein-like (PA) gene, complete sequence.

gi|54301549|AY664766| Influenza A virus (A/chicken/Hong Kong/WF126/03(H9N2)) polymerase acid protein (PA) gene, partial coding sequence.

gi|54301551|AY664767| Influenza A virus (A/pigeon/Hong Kong/WF53/03(H9N2)) polymerase acid protein (PA) gene, partial coding sequence.

gi|54301553|AY664768| Influenza A virus (A/pheasant/Hong Kong/WF54/03(H9N2)) polymerase acid protein (PA) gene, partial coding sequence.

gi|54301555|AY664769| Influenza A virus (A/Grizzly chicken/Hong Kong/NT184/03(H9N2)) polymerase acid protein (PA) gene, partial coding sequence.

gi|54301557|AY664770| Influenza A virus (A/chicken/Hong Kong/WF120/03(H9N2)) polymerase acid protein (PA) gene, partial coding sequence.

gi|54301559|AY664771| Influenza A virus (A/chicken/Hong Kong/NT366/03(H9N2)) polymerase acid protein (PA) gene, partial coding sequence.

gi|54301561|AY664772| Influenza A virus (A/chicken/Hong Kong/SSP418/03(H9N2)) polymerase acid protein (PA) gene, partial coding sequence.

gi|54301563|AY664773| Influenza A virus (A/chicken/Hong Kong/YU427/03(H9N2)) polymerase acid protein (PA) gene, partial coding sequence.

gi|55793682|AY800238| Influenza A virus (A/chicken/Korea/S1/2003(H9N2)) polymerase acid protein (PA) gene, complete coding sequence.

gi|58429718|AY862678| Influenza A virus (A/silky chicken/Korea/S3/03(H9N2)) PA (PA) gene, partial coding sequence.

gi|58429720|AY862679| Influenza A virus (A/chicken/Korea/S4/03(H9N2)) PA(PA) gene, partial coding sequence.

gi|58429722|AY862680| Influenza A virus (A/Chicken/Korea/S5/03(H9N2)) PA(PA) gene, complete coding sequence.

gi|58429724|AY862681| Influenza A virus (A/Chicken/Korea/S12/03(H9N2)) PA(PA) gene, partial coding sequence.

gi|58429726|AY862682| Influenza A virus (A/duck/Korea/S13/03(H9N2)) PA(PA) gene, partial coding sequence.

gi|58429728|AY862683| Influenza A virus (A/pigeon/Korea/S14/03(H9N2)) PA(PA) gene, partial coding sequence.

gi|58429730|AY862684| Influenza A virus (A/Chicken/Korea/S15/03(H9N2)) PA(PA) gene, partial coding sequence.

gi|58429732|AY862685| Influenza A virus (A/Chicken/Korea/S16/03(H9N2)) PA(PA) gene, partial coding sequence.

gi|58429734|AY862686| Influenza A virus (A/Chicken/Korea/S18/03(H9N2)) PA(PA) gene, partial coding sequence.

gi|5732356|AF156444| Influenza A virus (A/chicken/Hong Kong/G9/97(H9N2)) fragment 3 polymerase (PA) gene, partial coding sequence.

gi|5732358|AF156445| Influenza A virus (A/chicken/Hong Kong/G23/97(H9N2)) segment 3 polymerase (PA) gene, complete coding sequence.

gi|5732360|AF156446| Influenza A virus (A/pigeon/Hong Kong/Y233/97(H9N2)) fragment 3 polymerase (PA) gene, partial coding sequence.

gi|5732362|AF156447| Influenza A virus (A/duck/Hong Kong/Y280/97(H9N2)) segment 3 polymerase (PA) gene, partial coding sequence.

gi|5732364|AF156448| Influenza A virus (A/duck/Hong Kong/Y439/97(H9N2)) fragment 3 polymerase (PA) gene, partial coding sequence.

gi|5732366|AF156449| Influenza A virus (A/quail/Hong Kong/G1/97(H9N2)) fragment 3 polymerase (PA) gene, partial coding sequence.

gi|5732368|AF156450| Influenza A virus (A/chicken/Hong Kong/739/94(H9N2)) segment 3 polymerase (PA) gene, partial coding sequence.

gi|5732370|AF156451| Influenza A virus (A/quail/Hong Kong/AF157/92(H9N2)) fragment 3 polymerase (PA) gene, partial coding sequence.

gi|5732372|AF156452| Influenza A virus (A/chicken/Beijing/1/94(H9N2)) fragment 3 polymerase (PA) gene, partial coding sequence.

gi|5732374|AF156453| Influenza A virus (A/chicken/Korea/38349-p96323/96(H9N2)) segment 3 polymerase (PA) gene, partial coding sequence.

gi|5732376|AF156454| Influenza A virus (A/chicken/Korea/25232-96006/96(H9N2)) fragment 3 polymerase (PA) gene, partial coding sequence.

gi|5732378|AF156455| Influenza A virus (A/Shorebird/Delaware/9/96 (H9N2)) segment 3 polymerase (PA) gene, partial coding sequence.

gi|5732380|AF156456| Influenza A virus (A/quail/Arkansas/29209-1/93(H9N2)) fragment 3 polymerase (PA) gene, partial coding sequence.

gi|5732382|AF156457| Influenza A virus (A/Turkey/California/189/66(H9N2)) fragment 3 polymerase (PA) gene, partial coding sequence.

gi|12060671|AF222642| Influenza A virus (A/quail/Hong Kong/A17/99(H9N2)) fragment 3 PA (PA) gene, partial coding sequence.

gi|12060673|AF222643| Influenza A virus (A/pigeon/Hong Kong/FY6/99(H9N2)) fragment 3 PA(PA) gene, partial coding sequence.

gi|12060675|AF222644| Influenza A virus (A/chicken/Hong Kong/NT16/99(H9N2)) fragment 3 PA (PA) gene, partial coding sequence.

gi|12060677|AF222645| Influenza A virus (A/quail/Hong Kong/SSP10/99(H9N2)) fragment 3 PA (PA) gene, partial coding sequence.

gi|12060679|AF222646| Influenza A virus (A/pheasant/Hong Kong/SSP11/99(H9N2)) fragment 3 PA(PA) gene, partial coding sequence.

gi|12060681|AF222647| Influenza A virus (A/chicken/Hong Kong/FY20/99(H9N2)) fragment 3 PA (PA) gene, partial coding sequence.

gi|12060683|AF222648| Influenza A virus (A/chicken/Hong Kong/KC12/99(H9N2)) fragment 3 PA(PA) gene, partial coding sequence.

gi|12060685|AF222649| Influenza A virus (A/quail/Hong Kong/NT28/99(H9N2)) fragment 3 PA (PA) gene, partial coding sequence.

gi|12060687|AF222650| Influenza A virus (A/chicken/Hong Kong/SF2/99(H9N2)) fragment 3 PA(PA) gene, partial coding sequence.

gi|12060689|AF222651| Influenza A virus (A/Silkie/Hong Kong/SF44/99(H9N2)) fragment 3 PA(PA) gene, partial coding sequence.

Claims (23)

1. the iRNA reagent that comprises sense strand and antisense strand, wherein said sense strand comprises at least 15 successive Nucleotide, described successive Nucleotide and the reagent that provides of table 1A-1H are numbered no more than 1,2 or 3 of the different IPs thuja acid of sense strand sequence of arbitrary reagent of AL-DP-2241-AL-DP-8631, wherein said antisense strand comprises at least 15 successive Nucleotide, and the reagent that described successive Nucleotide and table provide among the 1A-1H is numbered no more than 1,2 or 3 of the different IPs thuja acid of antisense sequences of arbitrary reagent of AL-DP-2241-AL-DP-8631.

2. the iRNA reagent that comprises sense strand and antisense strand, wherein said sense strand comprises at least 15 successive Nucleotide, described successive Nucleotide and the reagent that provides of table 1A-1H are numbered the different IPs thuja acid no more than 1 of sense strand sequence of arbitrary reagent of AL-DP-2241-AL-DP-8631,2 or 3, the reagent that wherein said antisense strand comprises table to be provided among the 1A-1H is numbered at least 15 successive Nucleotide of antisense sequences of arbitrary reagent of AL-DP-2241-AL-DP-8631, and described iRNA reagent makes the target gene expression of expressing in the Cos-7 cell of target gene separately through transformation separately reduce by 20% than the expression in the cell of iRNA reagent incubation not, 30%, 40%, 50%, 60%, more than 70% or 80%.

3. the iRNA reagent that comprises sense strand and antisense strand, sense strand and antisense strand all comprise at least 16,17 or 18 nucleotide sequences, to be numbered the sequence of arbitrary reagent of AL-DP-2241-AL-DP-8631 identical with reagent that table provides among the 1A-1H basically for described sequence, different is that each chain has no more than 1,2 or 3 Nucleotide to be replaced (for example adenosine is replaced by uridylic) by other Nucleotide, forms the ability that reduces the amount of the influenza A plaque that forms in the cell in the detection but still kept basically in plaque.

4. each described iRNA reagent in the claim 1-3, the length of wherein said sense-rna chain are 30 Nucleotide or still less, and the length in the duplex zone of described iRNA reagent is 15-30 nucleotide pairs.

5. each described iRNA reagent in the claim 1-3, it comprises can make the modification that stability increases in biological sample of described iRNA reagent.

6. each described iRNA reagent in the claim 1-3, it comprises thiophosphatephosphorothioate or the 2 ' Nucleotide of modifying.

7. each described iRNA reagent in the claim 1-3, it comprises at least one 5 '-uridine-VITAMIN B4-3 ' (5 '-ua-3 ') dinucleotides, and wherein said uridine is the 2 ' Nucleotide of modifying; At least one 5 '-uridine-guanine-3 ' (5 '-ug-3 ') dinucleotides, wherein said 5 '-uridine is the 2 ' Nucleotide of modifying; At least one 5 '-cytidine-VITAMIN B4-3 ' (5 '-ca-3 ') dinucleotides, wherein said 5 '-cytidine is the 2 ' Nucleotide of modifying; Or at least one 5 '-uridine-uridine-3 ' (5 '-uu-3 ') dinucleotides, wherein said 5 '-uridine is the 2 ' Nucleotide of modifying.

8. claim 6 or 7 described iRNA reagent, wherein said 2 ' modifies the group that is selected from following composition: 2 '-deoxidation, 2 '-deoxidation-2 '-fluorine, 2 '-O-methyl, 2 '-the O-methoxy ethyl (2 '-O-MOE), 2 '-the O-aminopropyl (2 '-O-AP), 2 '-the O-dimethyl aminoethyl (2 '-O-DMAOE), 2 '-the O-dimethylaminopropyl (2 '-O-DMAP), 2 '-O-dimethyl aminoethyl oxygen ethyl (2 '-O-DMAEOE) and 2 '-O-N-methyl kharophen (2 '-O-NMA).

9. each described iRNA reagent of claim 1-3, it comprises the Nucleotide overhang with 1-4 unpaired nucleotides.

10. the described iRNA reagent of claim 9, the overhang of wherein said Nucleotide has 2 or 3 unpaired Nucleotide.

11. the described iRNA reagent of claim 9, wherein said Nucleotide overhang is at 3 ' end of described iRNA reagent antisense strand.

12. each described iRNA reagent of claim 1-3, it comprises cholesterol moiety.

13. the described iRNA reagent of claim 12, wherein said cholesterol moiety combines with 3 ' end of described iRNA reagent sense strand.

14. each described iRNA reagent of claim 1-3, wherein said iRNA reagent target is in being absorbed by pneumonocyte.

15. each described iRNA reagent of claim 1-3, wherein said iRNA reagent comprises at least a non-natural base.

16. the described iRNA reagent of claim 15, wherein said non-natural base are difluoro toluene base, nitroindoline base, nitro-pyrrole base or nitroimidazole base.

17. the described iRNA reagent of claim 15, wherein said non-natural base is the difluoro toluene base.

18. the described iRNA reagent of claim 15 only wherein comprises in two oligonucleotide chains of double chain oligonucleotide one and contains the non-natural base.

19. the described iRNA reagent of claim 15, two oligonucleotide chains that wherein comprise double chain oligonucleotide contain the non-natural base respectively.

20. treat its pathologic process part is duplicated the experimenter who is mediated by influenza A virus method, wherein said iRNA pack contains sense strand and antisense strand, wherein said sense strand comprises at least 15 successive Nucleotide, described successive Nucleotide and the reagent that provides of table 1A-1H are numbered the different IPs thuja acid no more than 1 of sense strand sequence of arbitrary reagent of AL-DP-2241-AL-DP-8631,2 or 3, wherein said antisense strand comprises at least 15 successive Nucleotide, and the reagent that described successive Nucleotide and table provide among the 1A-1H is numbered the different IPs thuja acid no more than 1 of antisense strand sequence of arbitrary reagent of AL-DP-2241-AL-DP-8631,2 or 3.

21. method according to claim 20, wherein said iRNA reagent is used with the amount of duplicating that enough reduces influenza virus in experimenter's cell or tissue.

22. method according to claim 20, wherein said experimenter is the people.

23. pharmaceutical composition comprises:

A.) each described iRNA reagent in the claim 1-3;

B.) pharmaceutical acceptable carrier.

Images