CN1324820A - New human protein with the function of inhibiting cancer cell growth and its encoding sequence - Google Patents

Abstract

The invention discloses a novel human protein with tumor suppressor function, a polynucleotide encoding the polypeptide and a method for producing the polypeptide through recombinant technology. The invention also discloses a method for using the polypeptide to treat various diseases such as cancer and the like. The invention also discloses an antagonist against the polypeptide and its therapeutic effect. The invention also discloses the application of the polynucleotide encoding this novel human protein with tumor suppressor function.

Description

Novel human protein with the function of inhibiting the growth of cancer cells and its coding sequence

The invention belongs to the field of biotechnology, in particular, the invention relates to a new polynucleotide encoding a human protein with tumor suppressor function, and a polypeptide encoded by the polynucleotide. The present invention also relates to the use and preparation of such polynucleotides and polypeptides.

Human genomics research is currently a hot spot in the world. In addition to large-scale sequencing of human chromosomal DNA and expressed sequence sequencing (EST), there is still a lack of high-throughput methods for screening functional genes starting from function.

Cancer is one of the major diseases that endanger human health. In order to effectively treat and prevent tumors, people have paid more and more attention to gene therapy of tumors. Therefore, there is an urgent need in this field to develop and study human proteins with tumor suppressor functions and their agonists/inhibitors.

The object of the present invention is to provide a new class of human protein polypeptides with tumor suppressor function and fragments, analogs and derivatives thereof.

Another object of the present invention is to provide polynucleotides encoding these polypeptides.

Another object of the present invention is to provide methods for producing these polypeptides and uses of the polypeptides and coding sequences.

In the first aspect of the present invention, a novel isolated protein polypeptide with tumor suppressor function is provided, which comprises a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO: 2, SEQ ID NO: 5, SEQ ID NO :8, SEQ ID NO:11, SEQ ID NO:14, SEQ ID NO:17, SEQ ID NO:20, SEQ ID NO:23, SEQ ID NO:26; or its conservative variant polypeptide, or its active fragment , or its active derivatives.

Preferably, the polypeptide is a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:5, SEQ ID NO:8, SEQ ID NO:11, SEQ ID NO:14, SEQ ID NO:17, SEQ ID NO:20, SEQ ID NO:23, SEQ ID NO:26.

In a second aspect of the present invention there is provided an isolated polynucleotide comprising a nucleotide sequence having at least 85% identity to a nucleotide sequence selected from the group consisting of: (a) a polynucleotide encoding the above-mentioned protein polypeptide with a tumor suppressor function; (b) a polynucleotide complementary to the polynucleotide (a). Preferably, the polypeptide encoded by the polynucleotide has an amino acid sequence selected from the group consisting of SEQ ID NO: 2, SEQ ID NO: 5, SEQ ID NO: 8, SEQ ID NO: 11, SEQ ID NO: 14, SEQ ID NO: 14, SEQ ID NO: ID NO:17, SEQ ID NO:20, SEQ ID NO:23, SEQ ID NO:26. More preferably, the sequence of the polynucleotide is selected from the group consisting of SEQ ID NO:3, SEQ ID NO:6, SEQ ID NO:9, SEQ ID NO:12, SEQ ID NO:15, SEQ ID NO:18 , SEQ ID NO:21, SEQ ID NO:24, SEQ ID NO:27 coding region sequence or full-length sequence.

In the third aspect of the present invention, there are provided vectors containing the above-mentioned polynucleotides, and host cells transformed or transduced by the vectors or host cells directly transformed or transduced by the above-mentioned polynucleotides.

In the fourth aspect of the present invention, there is provided a method for preparing a polypeptide having a protein activity of a tumor suppressor function, the method comprising: (a) cultivating the above-mentioned transformed or transformed protein under conditions suitable for expressing a protein with a tumor suppressor function (b) isolating the polypeptide with protein activity of tumor suppressor function from the culture.

In the fifth aspect of the present invention, an antibody specifically binding to the above-mentioned protein polypeptide with tumor suppressor function is provided. Also provided is a nucleic acid molecule useful for detection, which contains consecutive 10-800 nucleotides of the above-mentioned polynucleotides.

In the sixth aspect of the present invention, a pharmaceutical composition is provided, which contains a safe and effective amount of the protein polypeptide with tumor suppressor function of the present invention and a pharmaceutically acceptable carrier. These pharmaceutical compositions can treat diseases such as cancer and abnormal proliferation of cells.

Other aspects of the invention will be apparent to those skilled in the art from the technical disclosure herein.

The invention adopts large-scale cDNA clones to transfect cancer cells, and on the basis of obtaining tumor-suppressing effects, it is proved to be new genes by sequencing, and further obtains full-length cDNA clones. The DNA transfection test proves that the protein with tumor suppressor function of the present invention has the effect of inhibiting the colony formation of cancer cells (liver cancer cells), and the inhibition rate is 50% or above.

As used herein, "isolated" means that the material is separated from its original environment (if the material is native, the original environment is the natural environment). For example, polynucleotides and polypeptides in the natural state in living cells are not isolated and purified, but the same polynucleotides or polypeptides are isolated and purified if they are separated from other substances that exist together in the natural state .

As used herein, "isolated protein or polypeptide with tumor suppressor function" means that the protein or polypeptide with tumor suppressor function does not substantially contain other proteins, lipids, carbohydrates or other substances associated with it in nature. Those skilled in the art can use standard protein purification techniques to purify proteins with tumor suppressor function. Substantially pure polypeptides yield a single major band on non-reducing polyacrylamide gels. The purity of the protein polypeptide with tumor suppressor function can be analyzed by amino acid sequence.

The polypeptide of the present invention can be a recombinant polypeptide, a natural polypeptide, a synthetic polypeptide, preferably a recombinant polypeptide. Polypeptides of the present invention may be naturally purified, or chemically synthesized, or produced using recombinant techniques from prokaryotic or eukaryotic hosts (eg, bacteria, yeast, higher plants, insect and mammalian cells). Depending on the host used in the recombinant production protocol, the polypeptides of the invention may be glycosylated, or may be non-glycosylated. Polypeptides of the invention may or may not include an initial methionine residue.

The present invention also includes fragments, derivatives and analogs of human proteins with tumor suppressor function. As used herein, the terms "fragment", "derivative" and "analogue" refer to a polypeptide that substantially maintains the same biological function or activity of the natural human protein with tumor suppressor function of the present invention. The polypeptide fragments, derivatives or analogs of the present invention may be (i) polypeptides having one or more conservative or non-conservative amino acid residues (preferably conservative amino acid residues) substituted, and such substituted amino acid residues It may or may not be encoded by the genetic code, or (ii) a polypeptide having a substituent group in one or more amino acid residues, or (iii) a mature polypeptide in combination with another compound (such as a compound that extends the half-life of the polypeptide, e.g. polyethylene glycol), or (iv) an additional amino acid sequence fused to the polypeptide sequence (such as a leader sequence or secretory sequence or a sequence used to purify the polypeptide or a proprotein sequence). Such fragments, derivatives and analogs are within the purview of those skilled in the art in light of the teachings herein.

A polynucleotide of the invention may be in the form of DNA or RNA. Forms of DNA include cDNA, genomic DNA or synthetic DNA. DNA can be single-stranded or double-stranded. DNA can be either the coding strand or the non-coding strand. Taking the PP4277 protein (in this application, the naming of the protein adopts its clone number) (in this application, the naming of the protein adopts its clone number) as an example, the sequence of the coding region encoding the mature polypeptide can be shown in SEQ ID NO:3 The coding region sequences of the same or degenerate variants. As used herein, "degenerate variant" in the present invention refers to a nucleic acid sequence that encodes a protein having SEQ ID NO:2, but differs from the sequence of the coding region shown in SEQ ID NO:3. Taking the PP4377 protein (in this application, the naming of the protein adopts its clone number) (in this application, the naming of the protein adopts its clone number) as an example, the sequence of the coding region encoding the mature polypeptide can be shown in SEQ ID NO: 6 The coding region sequences of the same or degenerate variants. As used herein, "degenerate variant" in the present invention refers to a nucleic acid sequence that encodes a protein having SEQ ID NO:5, but differs from the sequence of the coding region shown in SEQ ID NO:6. For other proteins with tumor suppressor function, it can be deduced by analogy. For other proteins with tumor suppressor function, it can be deduced by analogy.

A polynucleotide encoding a mature polypeptide includes: a coding sequence that encodes only the mature polypeptide; a coding sequence for the mature polypeptide and various additional coding sequences; a coding sequence for the mature polypeptide (and optionally additional coding sequences) and non-coding sequences.

The term "polynucleotide encoding a polypeptide" may include a polynucleotide encoding the polypeptide, or may also include additional coding and/or non-coding sequences.

The present invention also relates to variants of the above-mentioned polynucleotides, which encode polypeptides or polypeptide fragments, analogs and derivatives having the same amino acid sequence as the present invention. Variants of this polynucleotide may be naturally occurring allelic variants or non-naturally occurring variants. These nucleotide variants include substitution variants, deletion variants and insertion variants. As known in the art, an allelic variant is an alternative form of a polynucleotide which may be a substitution, deletion or insertion of one or more nucleotides without substantially altering the function of the polypeptide it encodes .

The present invention also relates to polynucleotides that hybridize to the above-mentioned sequences and have at least 50%, preferably at least 70%, more preferably at least 80% identity between the two sequences. The invention particularly relates to polynucleotides which are hybridizable under stringent conditions to the polynucleotides of the invention. In the present invention, "stringent conditions" refer to: (1) hybridization and elution at lower ionic strength and higher temperature, such as 0.2×SSC, 0.1% SDS, 60°C; or (2) hybridization with There are denaturing agents, such as 50% (v/v) formamide, 0.1% calf serum/0.1% Ficoll, 42°C, etc.; or (3) only the identity between the two sequences is at least 95%, more Preferably hybridization occurs above 97%. Moreover, the polypeptide encoded by the hybridizable polynucleotide has the same biological function and activity as the mature polypeptide shown in SEQ ID NO:2.

The present invention also relates to nucleic acid fragments that hybridize to the above-mentioned sequences. As used herein, a "nucleic acid fragment" is at least 15 nucleotides in length, preferably at least 30 nucleotides in length, more preferably at least 50 nucleotides in length, most preferably at least 100 nucleotides in length. Nucleic acid fragments can be used in nucleic acid amplification techniques (such as PCR) to identify and/or isolate polynucleotides encoding proteins with tumor suppressor function.

The polypeptides and polynucleotides of the invention are preferably provided in isolated form, more preferably purified to homogeneity.

The DNA sequences of the present invention can be obtained in several ways. For example, DNA is isolated using hybridization techniques well known in the art. These techniques include, but are not limited to: 1) hybridization of probes to genomic or cDNA libraries to detect homologous nucleotide sequences, and 2) antibody screening of expression libraries to detect cloned DNA fragments with common structural features .

The production of specific DNA fragment sequences encoding proteins with tumor suppressor function can also be obtained by the following methods: 1) isolation of double-stranded DNA sequences from genomic DNA; 2) chemical synthesis of DNA sequences to obtain double-stranded DNA of desired polypeptides.

Of the methods mentioned above, isolating genomic DNA is the least commonly used. Direct chemical synthesis of DNA sequences is often the method of choice when the entire amino acid sequence of the desired polypeptide product is known. If the entire sequence of the desired amino acids is not known, direct chemical synthesis of the DNA sequence is not possible and the method of choice is isolation of the cDNA sequence. The standard method for isolating cDNA of interest is to isolate mRNA from donor cells that highly express the gene and perform reverse transcription to form a plasmid or phage cDNA library. There are many mature technologies for the method of extracting mRNA, and the kit is also available from commercial sources (Qiagene). It is also a common method to construct a cDNA library (Sambrook, et al., Molecular Cloning, A Laboratory Manual, Cold Spring Harbor Laboratory. New York, 1989). Commercially available cDNA libraries are also available, such as various cDNA libraries from the company Clontech. When combined with polymerase reaction technology, even minimal expression products can be cloned.

These cDNA libraries can be screened for the gene of the present invention by a conventional method. These methods include (but are not limited to): (1) DNA-DNA or DNA-RNA hybridization; (2) function appearance or loss of marker genes; (3) measuring the level of transcripts of proteins with tumor suppressor function; (4) ) Detection of protein products expressed by genes by immunological techniques or assays of biological activity. The above methods can be used alone or in combination with multiple methods.

In the (1) method, the probe used for hybridization is homologous to any part of the polynucleotide of the present invention, and its length is at least 15 nucleotides, preferably at least 30 nucleotides, more preferably At least 50 nucleotides, preferably at least 100 nucleotides. In addition, the length of the probe is usually within 2kb, preferably within 1kb. The probes used here are usually DNA sequences chemically synthesized based on the gene DNA sequence information of the present invention. The genes themselves or fragments of the present invention can of course be used as probes. DNA probes can be labeled with radioactive isotopes, luciferin or enzymes (such as alkaline phosphatase) and the like.

In the (4) method, immunological techniques such as Western blotting, radioimmunoprecipitation, and enzyme-linked immunosorbent assay (ELISA) can be used to detect the protein product expressed by the protein gene with tumor suppressor function.

A method of amplifying DNA/RNA using the PCR technique (Saiki, et al. Science 1985; 230:1350-1354) is preferably used to obtain the gene of the present invention. Especially when it is difficult to obtain full-length cDNA from the library, the RACE method (RACE-cDNA terminal rapid amplification method) can be preferably used, and the primers used for PCR can be appropriately selected according to the sequence information of the present invention disclosed herein, And can be synthesized by conventional methods. Amplified DNA/RNA fragments can be separated and purified by conventional methods such as by gel electrophoresis.

The determination of the nucleotide sequence of the gene of the present invention obtained as described above, or various DNA fragments, etc., can be performed by conventional methods such as the dideoxy chain termination method (Sanger et al. PNAS, 1977, 74:5463-5467). Such nucleotide sequence determination can also use commercial sequencing kits and the like. In order to obtain the full-length cDNA sequence, sequencing needs to be repeated. Sometimes it is necessary to determine the cDNA sequence of multiple clones before splicing into a full-length cDNA sequence.

The present invention also relates to a vector containing the polynucleotide of the present invention, a host cell produced by genetic engineering using the vector of the present invention or a protein coding sequence with tumor suppressor function, and a method for producing the polypeptide of the present invention by recombinant technology.

Through conventional recombinant DNA technology, the polynucleotide sequence of the present invention can be used to express or produce recombinant protein polypeptide with tumor suppressor function (Science, 1984; 224:1431). Generally speaking, there are the following steps:

(1). Transform or transduce suitable host cells with the polynucleotide (or variant) encoding the human protein with tumor suppressor function of the present invention, or with the recombinant expression vector containing the polynucleotide;

(2). host cells cultured in a suitable medium;

(3). Isolate and purify proteins from culture medium or cells.

In the present invention, the human protein polynucleotide sequence with tumor suppressor function can be inserted into the recombinant expression vector. The term "recombinant expression vector" refers to bacterial plasmid, phage, yeast plasmid, plant cell virus, mammalian cell virus such as adenovirus, retrovirus or other vectors well known in the art. Carriers suitable for use in the present invention include, but are not limited to: T7-based expression vectors expressed in bacteria (Rosenberg, et al. Gene, 1987, 56:125); pMSXND expression vectors expressed in mammalian cells (Lee and Nathans, J Bio Chem.263:3521, 1988) and vectors derived from baculovirus expressed in insect cells. In short, any plasmid and vector can be used as long as it can be replicated and stabilized in the host. An important feature of expression vectors is that they usually contain an origin of replication, a promoter, marker genes, and translational control elements.

Methods well known to those skilled in the art can be used to construct expression vectors containing human protein-coding DNA sequences with tumor suppressor function and appropriate transcription/translation control signals. These methods include in vitro recombinant DNA technology, DNA synthesis technology, in vivo recombination technology, etc. (Sambroook, et al. Molecular Cloning, a Laboratory Manual, cold Spring Harbor Laboratory. New York, 1989). Said DNA sequence can be operably linked to an appropriate promoter in the expression vector to direct mRNA synthesis. Representative examples of these promoters are: E. coli lac or trp promoter; lambda phage PL promoter; eukaryotic promoters include CMV immediate early promoter, HSV thymidine kinase promoter, early and late SV40 promoter, reverse LTRs of transcription viruses and other promoters known to control the expression of genes in prokaryotic or eukaryotic cells or their viruses. The expression vector also includes a ribosome binding site for translation initiation and a transcription terminator.

In addition, the expression vector preferably contains one or more selectable marker genes to provide phenotypic traits for selection of transformed host cells, such as dihydrofolate reductase for eukaryotic cell culture, neomycin resistance, and green Fluorescent protein (GFP), or tetracycline or ampicillin resistance for E. coli.

Vectors containing the above-mentioned appropriate DNA sequences and appropriate promoters or control sequences can be used to transform appropriate host cells so that they can express proteins.

The host cell may be a prokaryotic cell, such as a bacterial cell; or a lower eukaryotic cell, such as a yeast cell; or a higher eukaryotic cell, such as a mammalian cell. Representative examples are: Escherichia coli, Streptomyces spp; bacterial cells of Salmonella typhimurium; fungal cells such as yeast; plant cells; insect cells of Drosophila S2 or Sf9; animal cells of CHO, COS or Bowes melanoma cells, etc.

When the polynucleotide of the present invention is expressed in higher eukaryotic cells, if an enhancer sequence is inserted into the vector, the transcription will be enhanced. Enhancers are cis-acting elements of DNA, usually about 10 to 300 base pairs in length, that act on promoters to enhance gene transcription. Examples include the SV40 enhancer of 100 to 270 base pairs on the late side of the replication origin, the polyoma enhancer on the late side of the replication origin, and the adenovirus enhancer.

Those of ordinary skill in the art will know how to select appropriate vectors, promoters, enhancers and host cells.

Transformation of host cells with recombinant DNA can be performed using conventional techniques well known to those skilled in the art. When the host is a prokaryotic organism such as E. coli, competent cells capable of taking up DNA can be harvested after the exponential growth phase and treated with the _CaCl2 method using procedures well known in the art. An alternative is to use _MgCl2 . Transformation can also be performed by electroporation, if desired. When the host is eukaryotic, the following DNA transfection methods can be used: calcium phosphate co-precipitation method, conventional mechanical methods such as microinjection, electroporation, liposome packaging, etc.

The obtained transformant can be cultured by conventional methods to express the polypeptide encoded by the gene of the present invention. The medium used in the culture can be selected from various conventional media according to the host cells used. The culture is carried out under conditions suitable for the growth of the host cells. After the host cells have grown to an appropriate cell density, the selected promoter is induced by an appropriate method (such as temperature shift or chemical induction), and the cells are cultured for an additional period of time.

The recombinant polypeptide in the above method can be encapsulated inside the cell, outside the cell or expressed on the cell membrane or secreted outside the cell. The recombinant protein can be isolated and purified by various separation methods by taking advantage of its physical, chemical and other properties, if desired. These methods are well known to those skilled in the art. Examples of these methods include, but are not limited to: conventional refolding treatment, treatment with protein precipitating agents (salting out method), centrifugation, osmotic disruption, supertreatment, ultracentrifugation, molecular sieve chromatography (gel filtration), adsorption layer Analysis, ion exchange chromatography, high performance liquid chromatography (HPLC) and various other liquid chromatography techniques and combinations of these methods.

The recombinant human protein or polypeptide with tumor suppressor function has many uses. These uses include (but are not limited to): direct use as a drug to treat diseases caused by the hypofunction or loss of proteins with tumor suppressor functions, and screening for antibodies, polypeptides or other ligands that promote or resist the functions of proteins with tumor suppressor functions. body. For example, antibodies can be used to activate or inhibit the function of human proteins that function as tumor suppressors. Screening the polypeptide library with the expressed recombinant human protein with tumor suppressor function can be used to find therapeutically valuable polypeptide molecules that can inhibit or stimulate the function of the human protein with tumor suppressor function.

The invention also provides methods of screening drugs to identify agents that enhance (agonists) or repress (antagonists) human proteins with tumor suppressor function. Agonists enhance biological functions of human proteins with tumor suppressor functions such as stimulating cell proliferation, while antagonists prevent and treat disorders related to excessive cell proliferation, such as various cancers. For example, mammalian cells or membrane preparations expressing a human protein with tumor suppressor function can be cultured with the labeled human protein with tumor suppressor function in the presence of a drug. The ability of the drug to enhance or repress this interaction is then determined.

Antagonists of human proteins with tumor suppressor function include screened antibodies, compounds, receptor deletions and analogs. The antagonist of the human protein with tumor suppressor function can combine with the human protein with tumor suppressor function and eliminate its function, or inhibit the production of the human protein with tumor suppressor function, or combine with the active site of the polypeptide so that the polypeptide cannot perform biological functions. Antagonists of human proteins that function as tumor suppressors are useful for therapeutic use.

When screening compounds as antagonists, a protein with tumor suppressor function can be added to a bioanalytical assay, and whether the compound is an antagonist can be determined by measuring the effect of the compound on the interaction between the protein with tumor suppressor function and its receptor. Receptor deletions and analogs that function as antagonists can be screened in the same manner as described above for screening compounds.

The polypeptide of the present invention can be directly used in the treatment of diseases, for example, various malignant tumors, abnormal cell proliferation and the like.

The polypeptides of the present invention, and fragments, derivatives, analogs thereof or their cells can be used as antigens to produce antibodies. These antibodies can be polyclonal or monoclonal. Polyclonal antibodies can be obtained by injecting the polypeptide directly into animals. Techniques for preparing monoclonal antibodies include hybridoma technology, trioma technology, human B-cell hybridoma technology, EBV-hybridoma technology, and the like.

The polypeptides and antagonists of the present invention can be used in combination with suitable pharmaceutical carriers. These carriers can be water, dextrose, ethanol, salts, buffers, glycerol and combinations thereof. The composition contains safe and effective doses of polypeptides or antagonists as well as carriers and excipients that do not affect the drug effect. These compositions can be used as medicine for disease treatment.

The invention also provides kits or kits comprising one or more containers containing one or more ingredients of the pharmaceutical compositions of the invention. Along with these containers, there may be an indicative notice given by the governmental regulatory agency that manufactures, uses or sells the drug or biological product reflecting its approval for human administration by the governmental regulatory agency that manufactures, uses or sells the drug or biological product. In addition, the polypeptide of the present invention can be used in combination with other therapeutic compounds such as TNF and the like.

The pharmaceutical compositions may be administered in a convenient manner, such as by topical, intravenous, intraperitoneal, intramuscular, subcutaneous, intranasal or intradermal routes of administration. Proteins with tumor suppressor function are administered in amounts effective to treat and/or prevent the particular indication. The amount and dosage range of the protein with tumor suppressor function administered to a patient will depend on many factors, such as the mode of administration, the health condition of the person to be treated, and the judgment of the diagnosing physician.

Polynucleotides of human proteins with tumor suppressor function can also be used for various therapeutic purposes. Gene therapy technology can be used to treat abnormalities in cell proliferation, development or metabolism due to non-expression or abnormal/inactive expression of proteins with tumor suppressor functions. Recombinant gene therapy vectors (such as viral vectors) can be designed to express mutated proteins with tumor suppressor functions to inhibit the activity of endogenous proteins with tumor suppressor functions. For example, a mutated protein with tumor suppressor function may be a shortened protein with tumor suppressor function that lacks a signal transduction domain, and although it can bind to a downstream substrate, it lacks signal transduction activity. Therefore, the recombinant gene therapy vector can be used to treat diseases caused by abnormal expression or activity of proteins with tumor suppressor function. Expression vectors derived from viruses such as retroviruses, adenoviruses, adeno-associated viruses, herpes simplex viruses, and parvoviruses can be used to transfer protein genes with tumor suppressor functions into cells. The method for constructing a recombinant viral vector carrying a protein gene with a tumor suppressor function can be found in existing literature (Sambrook, et al.). In addition, the recombinant human protein gene with tumor suppressor function can be packaged into liposomes and transferred into cells.

Oligonucleotides (including antisense RNA and DNA) and ribozymes that inhibit human protein mRNA with tumor suppressor function are also within the scope of the invention. A ribozyme is an enzyme-like RNA molecule that can specifically decompose a specific RNA. Its mechanism of action is that the ribozyme molecule specifically hybridizes with a complementary target RNA to perform an endonucleic cut. Antisense RNA, DNA and ribozyme can be obtained by any existing RNA or DNA synthesis technology, such as solid-phase phosphoamide chemical synthesis of oligonucleotides, which has been widely used. Antisense RNA molecules can be obtained by in vitro or in vivo transcription of the DNA sequence encoding the RNA. This DNA sequence has been integrated into the vector downstream of the RNA polymerase promoter. In order to increase the stability of nucleic acid molecules, it can be modified in a variety of ways, such as increasing the sequence length on both sides, and the connection between ribonucleosides should use phosphothioester bonds or peptide bonds instead of phosphodiester bonds.

The methods for introducing polynucleotides into tissues or cells include: directly injecting polynucleotides into tissues in the body; or first introducing polynucleotides into cells in vitro through vectors (such as viruses, phages, or plasmids, etc.) , and then transplant the cells into the body, etc.

The polypeptide of the present invention can also be used for peptide spectrum analysis, for example, the polypeptide can be specifically cleaved physically, chemically or enzymatically, and subjected to one-dimensional, two-dimensional or three-dimensional gel electrophoresis analysis.

The invention also provides an antibody against the human protein antigenic determinant with tumor suppressor function. These antibodies include, but are not limited to: polyclonal antibodies, monoclonal antibodies, chimeric antibodies, single chain antibodies, Fab fragments and fragments produced by a Fab expression library.

Antibodies against human tumor suppressor proteins can be used in immunohistochemical techniques to detect human tumor suppressor proteins in biopsy specimens.

Monoclonal antibodies that bind to human proteins with tumor suppressor functions can also be labeled with radioactive isotopes, and injected into the body to track their location and distribution. This radiolabeled antibody can be used as a non-invasive diagnostic method for localization of tumor cells and judgment of metastasis.

The antibody of the present invention can be used to treat or prevent diseases related to the human protein with tumor suppressor function. Administration of appropriate doses of antibodies can stimulate or block the production or activity of human proteins with tumor suppressor functions.

Antibodies can also be used to design immunotoxins that target a particular site in the body. For example, high-affinity monoclonal antibodies to human proteins with tumor suppressor functions can be covalently combined with bacterial or plant toxins (such as diphtheria toxin, ricin, rhododine, etc.). A common method is to use a sulfhydryl cross-linking agent such as SPDP to attack the amino group of the antibody, and bind the toxin to the antibody through the exchange of disulfide bonds. This hybrid antibody can be used to kill human proteins with tumor suppressor functions. cell.

The production of polyclonal antibodies can be used to immunize animals, such as rabbits, mice, rats, etc., with human proteins or polypeptides with tumor suppressor functions. Various adjuvants can be used to enhance the immune response, including but not limited to Freund's adjuvant and the like.

Human protein monoclonal antibodies with tumor suppressor function can be produced by hybridoma technology (Kohler and Milstein. Nature, 1975, 256:495-497). Chimeric antibodies combining human constant regions and non-human variable regions can be produced using existing techniques (Morrison et al, PNAS, 1985, 81:6851). The existing technology for producing single-chain antibodies (U.S. Pat No. 4946778) can also be used to produce single-chain antibodies against human proteins with tumor suppressor functions.

Polypeptide molecules capable of binding to human proteins with tumor suppressor functions can be obtained by screening random polypeptide libraries composed of various possible combinations of amino acids bound to solid phases. During screening, human protein molecules with tumor suppressor function must be labeled.

The invention also relates to a diagnostic test method for quantitative and localized detection of the human protein level with tumor suppressor function. These assays are well known in the art and include FISH assays and radioimmunoassays. The level of human protein with tumor suppressor function detected in the test can be used to explain the importance of human protein with tumor suppressor function in various diseases and to diagnose diseases in which the protein with tumor suppressor function plays a role.

The polynucleotide of the protein with tumor suppressor function can be used for the diagnosis and treatment of diseases related to the protein with tumor suppressor function. In terms of diagnosis, the polynucleotide of the protein with tumor suppressor function can be used to detect the expression of the protein with tumor suppressor function or the abnormal expression of the protein with tumor suppressor function in a disease state. For example, DNA sequences of proteins with tumor suppressor function can be used for hybridization of biopsy specimens to determine abnormal expression of proteins with tumor suppressor function. Hybridization techniques include Southern blotting, Northern blotting, in situ hybridization, and the like. These technical methods are all open and mature technologies, and relevant kits are available from commercial sources. Part or all of the polynucleotides of the present invention can be used as probes to be immobilized on microarrays (Microarray) or DNA chips (also known as "gene chips") for analysis of differential expression of genes in tissues and gene diagnosis. RNA-polymerase chain reaction (RT-PCR) in vitro amplification with specific primers of the protein with tumor suppressor function can also detect the transcription product of the protein with tumor suppressor function.

The detection of the mutation of the protein gene with tumor suppressor function can also be used to diagnose the diseases related to the protein with tumor suppressor function. The form of protein mutation with tumor suppressor function includes point mutation, translocation, deletion, recombination and any other abnormality compared with the normal wild type protein DNA sequence with tumor suppressor function. Mutations can be detected using established techniques such as Southern blotting, DNA sequence analysis, PCR and in situ hybridization. In addition, mutations may affect protein expression, so Northern blotting and Western blotting can be used to indirectly determine whether a gene has a mutation.

The sequences of the invention are also valuable for chromosome identification. The sequence will be specific for a particular location on a human chromosome and can hybridize thereto. Currently, there is a need to identify the specific site of each gene on the chromosome. Currently, only a few chromosomal markers based on actual sequence data (repeat polymorphisms) are available to mark chromosomal positions. According to the present invention, in order to associate these sequences with disease-related genes, an important first step is to locate these DNA sequences on chromosomes.

In short, PCR primers (preferably 15-35bp) are prepared according to the cDNA, and the sequence can be positioned on the chromosome. These primers were then used for PCR screening of somatic heterozygous cells containing individual human chromosomes. Only those cells heterozygous for the human gene corresponding to the primer will produce an amplified fragment.

The PCR mapping method of somatic heterozygous cells is a quick method to locate DNA to specific chromosomes. Using the oligonucleotide primers of the present invention, sublocalization can be achieved using a set of fragments from a specific chromosome or a large number of genomic clones by a similar method. Other similar strategies that can be used for chromosome mapping include in situ hybridization, chromosome prescreening by flow sorting with markers, and hybridization preselection to construct chromosome-specific cDNA libraries.

Fluorescence in situ hybridization (FISH) of cDNA clones to metaphase chromosomes allows precise chromosomal mapping in a single step. For a review of this technique, see Verma et al., Human Chromosomes: a Manual of Basic Techniques, Pergamon Press, New York (1988).

Once a sequence has been mapped to an exact chromosomal location, the physical location of the sequence on the chromosome can be correlated with gene map data. These data can be found, eg, in V. Mckusick, Mendelian Inheritance in Man (available online through Johns Hopkins University Welch Medical Library). Linkage analysis can then be used to determine the relationship between the gene and the disease that has been mapped to the chromosomal region.

Next, the cDNA or genome sequence differences between affected and non-affected individuals need to be determined. If a mutation is observed in some or all of the affected individuals but not in any normal individual, the mutation may be the cause of the disease. Comparing affected and unaffected individuals usually involves first looking for structural changes in chromosomes, such as deletions or translocations that are visible at the chromosomal level or detectable with cDNA sequence-based PCR. Based on the resolution capabilities of current physical mapping and gene mapping techniques, the cDNA that is pinpointed to a disease-associated chromosomal region can be one of 50 to 500 potential disease-causing genes (assuming 1 megabase mapping resolution capacity and each 20kb corresponds to a gene).

The protein nucleotide full-length sequence or its fragments with tumor suppressor function of the present invention can usually be obtained by PCR amplification method, recombination method or artificial synthesis method. For the PCR amplification method, primers can be designed according to the relevant nucleotide sequences disclosed in the present invention, especially the open reading frame sequence, and the cDNA prepared by a commercially available cDNA library or a conventional method known to those skilled in the art can be used. The library is used as a template to amplify related sequences. When the sequence is long, it is often necessary to carry out two or more PCR amplifications, and then splice together the amplified fragments in the correct order.

Once the relevant sequences are obtained, recombinant methods can be used to obtain the relevant sequences in large quantities. Usually, it is cloned into a vector, then transformed into a cell, and then the relevant sequence is isolated from the proliferated host cell by conventional methods.

In addition, related sequences can also be synthesized by artificial synthesis, especially when the fragment length is relatively short. Often, fragments with very long sequences are obtained by synthesizing multiple small fragments and then ligating them.

At present, the DNA sequence encoding the protein of the present invention (or its fragments, or its derivatives) can be completely chemically synthesized. This DNA sequence can then be introduced into various DNA molecules (such as vectors) and cells known in the art. In addition, mutations can also be introduced into the protein sequences of the invention by chemical synthesis.

In addition, since the protein with tumor suppressor function of the present invention has a natural amino acid sequence derived from humans, it is expected to have higher activity and/or lower Low side effects (eg, less or no immunogenicity in humans).

Below in conjunction with specific embodiment, further illustrate the present invention. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental method that does not indicate specific conditions in the following examples, usually according to conventional conditions such as Sambrook et al., molecular cloning: the conditions described in the laboratory manual (New York: Cold Spring Harbor Laboratory Press, 1989), or according to the manufacturer suggested conditions.

Example 1: Obtaining of cDNA gene and its inhibitory effect on cancer cell clone formation

PP4277, PP4377, PP4522, PP4583, PP4664, PP4748, PP4762, PP5242, PP5656 were obtained by constructing human placenta cDNA library by conventional methods. Placental tissues at 3, 6, and 10 months old were taken, and total RNA was extracted with Trizol reagent (GIBCO BRL Company) according to the manufacturer's instructions, and mRNA was extracted with an mRNA purification kit (Pharmacia Company). The cDNA library of the above mRNA was constructed with the pCMV-script TMXR cDNA library construction kit (Stratagene). The reverse transcriptase was changed to MMLV-RT-Superscript II (GIBCO BRL), and the reverse transcription reaction was carried out at 42°C. XL10-Gold competent cells were transformed, and a cDNA library with a titer of 1×10 ⁶ cfu/μg cDNA was obtained. In the first round, cDNA clones were randomly selected, and then high-abundance cDNA clones and cDNA clones that had been proven to inhibit the growth of cancer cells were used as probes to hybridize and screen the cDNA library to pick weakly positive and negative clones. Plasmid DNA was extracted with Qiagen 96-well plate plasmid extraction kit according to the manufacturer's instructions. Plasmid DNA and empty vector were transfected into liver cancer cell line 7721 at the same time. After 100 ng of DNA was precipitated and dried by alcohol, 6 μl of H ₂ O was added to dissolve it and wait for transfection. Add 0.74 μl liposome and 9.3 μl serum-free medium to each DNA sample, mix well, and place at room temperature for 10 minutes. Add 150 μl of serum-free culture medium to each tube, add evenly to 7721 cells grown in 96-well plates in 3 wells, place at 37°C for 2 hours, add 50 μl of serum-free culture medium to each well, and keep at 37°C for 24 hours. Change 100 μl of whole culture solution per well, 37°C for 24 hours, change 100 μl of whole culture solution containing G418, 37°C for 24 to 48 hours, change the culture solution with different concentrations of G418 while observing. After about 2 to 3 times, until the microscopic examination of the cells showed colony formation, count them. It was found that the above clones can inhibit the formation of cell clones, and the results are shown in the table below. Cloning formation of cDNA clone transfected cells (7721) cDNA clone name Number of cDNA clones (three replicates) Number of empty vector clones (three replicates) PP4277 0 0 4 45 37 30 PP4377 1 16 4 33 12 16 PP4522 0 0 0 twenty three twenty one 14 PP4583 0 0 0 twenty three twenty one 14 PP4664 7 6 1 32 34 29 PP4748 0 2 1 twenty one 14 18 PP4762 4 11 0 32 29 43 PP5242 3 4 3 16 13 9 PP5656 0 0 0 35 39 32

The dideoxy termination method was used for the cDNA clone, and the nucleotide sequence of nearly 500 bp at one end was determined on an ABI377 DNA automatic sequencer. After analysis, it was determined to be a new gene clone, and the other end was sequenced. If the full-length cDNA sequence has not yet been obtained, then design primers and perform sequencing again until the full-length sequence (SEQ ID NO: 1, 4, 7, 10, 13, 16, 19, 22, 24) is obtained.

Embodiment 2: PCR obtains gene clone from placenta cDNA:

Placenta tissues at 3, 6, and 10 months old were taken, and total RNA was extracted with Trizol reagent (GIBCO BRL company) according to the manufacturer's instructions, and mRNA was extracted with mRNA purification kit (pharmacia company). Using MMLV-RT-Superscript II (GIBCO BRL) and reverse transcriptase at 42 ° C for reverse transcription reaction, to obtain placental cDNA. Utilize the transmutation primers of each gene (as shown in the table below), and perform 3'1 cycles at 97°C. PCR amplification was performed at 94°C 30″, 60°C 30″, 72°C 1′, 35 cycles, and 72°C 10′ 1 cycle to obtain amplification products of each gene. The amplified product was verified by sequencing and was consistent with the sequence measured in Example 1, and then the amplified product was transferred into host cells by conventional techniques to obtain the recombinant protein.

gene specific primer sequence clone name Specific primer 1 (5'→3') Specific primer 2 (5'→3') PP4277 TCACGGTCCCACTCAAGTCCTGC TCTGGACACAGAACGAGCAGCCC PP4377 GGCCCTGGGTTCACCTGATTGAG GAGCCCTCCTCCTAGGGCTGGAA PP4522 TCCCTGTCCCTGCTTCTCTTCGG AAGCAGAGGGCGAAAGCTGCAAG PP4583 GCGCGTCACCAGAGTCGTTTCTC GGTCTTTCCACTTCGGGCTTCTG PP4664 CAACGCGGACTTCATCATCCCTG CCACCTGATGCAGGCCAACCATA PP4748 CGACGCGCTGTCTGGAGTCGTAG TCTGCTGACATGAGGACCTGGGG PP4762 TGAATTGGCGCTATGTCTGGGGA TCAAGCACTTGGCTTCTGCTGCC PP5242 GGCTGGGTCCCACCAGTGACAAG CGACCTGCAGACGGGACATGACT PP5656 AGAGCTCTTTGTGTGCCCCAGCC AAGACCCCACCCCACAGATGCTC Example 3: cDNA clone sequence analysis 1. PP4277蛋白A：核苷酸序列(SEQ ID NO:1)长度：1915bpCACTAAGGCG CAAGGCAGCT GCACATCCAC GGCCAGCCTC GTTGTCTTCA GGGATTTGTT 60CAAAGTTCAT TCTGAATTGT AAGGTAACCA CTGCAGAACA CGAGCCTTTC CTGGGTATGT 120GTGGTTATTT CTGCCCTTCA CGGTCCCACT CAAGTCCTGC CTCCCTCAGG GAGACCTCCC 180AGACCTCACC AAGCCGTGGG CACGTGGACC AGAAGCCGTG GACCCCAGAG GCTCATTTAG 240GGATCTGCTT TCTGAACCCA GGTCCTAACA TCTGCACTGC AGCGAGCATT TCTGGTGCCT 300CAAGTGAGTC ACTCCAACTT CCAGCTCACT GAGAGGCGAT GGAGCACTGA GTGCCAGCAT 360CTTCCAGCAC CAGAGGAGCT GACTCTGAAT AGACAGCTCG GCAGCGGCCT CAGCACAGAG 420CAGGGGAGCC ACCAGCCACA GGCAGCCACT GAGCCCTTGG CATGTAACCC GGTAACCCAT 480GCAACAGAAG ACCTGAGCGT CCAGTGTTGC TAGCTTGAAG TAACTTCAGT GGTGTTCCCC 540AGCGTTTTTC TCATCCCCCT CCCTTCCCTT CCCCTACAGC TTTCTTCCTC TTTCCCCTCC 600TTCTGAGTCT TTTGTCCCCA GTCCTGGGAA CCTTTTTTCT GACTTGGTGT TTTCTCCTGA 660ATGCCTCCTT CCTGGCCCCC TAATCAGAAA TGCAGAAGCC ACCTGCCCCA CACAGGGGGC 720CGCTTTCCCT GCATCTGGCA CTCACCAGGA CAGTCATAGA ACTGGCCTTG GGTGCTTGGG 780GAGAGTGGAG AGGTCTGGGG GGCATCCACT GGCCTCAGCC TCAGAGCTTC TCATCAGTAA 840CCCAGGAGGC AAAGGCCCAG CTCGAGATTA GCCCAGGCCA CTGTATCTGT GCCAAGACCT 900ACAGCTGTCC CTGCAGGGCG TGCATCTCCT GCAGCCCAGG CTATCACAAT GGTGTCCCTT 960TTGGTGGGGA TGTGAGGCCT GCTGCCTGCC TGGGAAGAAG CTGACAGGAT TGTTGGTGCT 1020GTGGGGAGAA GCTGGTGTAA CTGAGACCCA TGAGGGAGGC CCTTTGTGGG TCTGTGGCTG 1080GGAGGAGCTC TGGGGAAGAT GGGCCTTGAG AAATCCCAGC TTAGCACCTT TGACAGTTGC 1140TCCACTGAGT CACCCTAGAC TGCAATCTCA CAGATGTTCT AACCACCCTC TCCCTGGGGA 1200GCCCTTGGGG GTGCTGCAAG CCCAGTCCTC ACTCACTGGC TTTGTGCTTC TCTTCCCGAC 1260TTCCTTTTTT TGCTCCTGTT GAGAAATGTG GGTCCAGGGC TGCGTGCTCC AGTGTGCGGA 1320AGGAAGGGCA GCGTGCTCTT TGGACTGAGC TGGCTCAAGG GAAGCTGGGT CTGAGGGGAG 1380ACTGACCTCG ACCCTTCTTA AATGAAACAC CTTTTCCCTG CGTCTCAGTC ACCAGTTACC 1440TGGCCTGATC ACTGTGTGCC ACACCCCATC ACATCCATCC CCACAAGAAT GCCAGGAGGA 1500GGGGTCATTG GCCTTTTTGC CAGCACAAAC CTCTGGAGGC CAGCTCCCAA GACCCGCCAG 1560GCACACCTGC CAGAGTGCCT TAACTTGGGA TGTCTGCAAA CCTCTGTCCA TGGCCGAACC 1620CACCCCACTG CCTGTTACTG TAAATAAAGG TCTATTAGCA TGCCACCATG CCCCTCCTCC 1680GATGCATACA GTGCCCAGCT CAGCACCTGG TGCAAAGTAA GCACAATCTG GCTTCCAGCC 1740ATCACTGCTG TTGCTACTAT CACAAATATT AGGCCCCTTC TAGAATGCAA AGATAACCCA 1800GGGCTGCTCG TTCTGTGTCC AGATCACAGA GTTTTTCTTT TTATCCTTAT CAACTTTATT 1860GAGGGATAAT TTACAGGCAA TAAATCATAT CCATTTAAAA AAAAAAAAAA AAAAA 1915B：氨基酸序列(SEQ ID NO:2)长度：104个氨基酸1 MQKPPAPHRG PLSLHLALTR TVIELALGAW GEWRGLGGIH WPQPQSFSSV TQEAKAQLEI 61 SPGHCICAKT YSCPCRACIS CSPGYHNGVP FGGDVRPAAC LGRSC． Nucleotide and amino acid combination sequence (SEQ ID NO: 3) Clone number: PP4277 Start codon: 690 ATG Stop codon: 1004 TGA Protein molecular weight: 110761 CA CTA AGG CGC AAG GCA GCT GCA CAT CCA CGG CCA GCC TCG TTG TCT 4748 TCA GGG ATT TGT TCA AAG TTC ATT CTG AAT TGT AAG GTA ACC ACT GCA 9596 GAA CAC GAG CCT TTC CTG GGT ATG TGT GGT TAT TTC TGC CCT TCA CGG 143144 TCC CAC TCA AGT CCT GCC TCC CTC AGG GAG ACC TCC CAG ACC TCA CCA 191192 AGC CGT GGG CAC GTG GAC CAG AAG CCG TGG ACC CCA GAG GCT CAT TTA 239240 GGG ATC TGC TTT CTG AAC CCA GGT CCT AAC ATC TGC ACT GCA GCG AGC 287288 ATT TCT GGT GCC TCA AGT GAG TCA CTC CAA CTT CCA GCT CAC TGA GAG 335336 GCG ATG GAG CAC TGA GTG CCA GCA TCT TCC AGC ACC AGA GGA GCT GAC 383384 TCT GAA TAG ACA GCT CGG CAG CGG CCT CAG CAC AGA GCA GGG GAG CCA 431432 CCA GCC ACA GGC AGC CAC TGA GCC CTT GGC ATG TAA CCC GGT AAC CCA 479480 TGC AAC AGA AGA CCT GAG CGT CCA GTG TTG CTA GCT TGA AGT AAC TTC 527528 AGT GGT GTT CCC CAG CGT TTT TCT CAT CCC CCT CCC TTC CCT TCC CCT 575576 ACA GCT TTC TTC CTC TTT CCC CTC CTT CTG AGT CTT TTG TCC CCA GTC 623624 CTG GGA ACC TTT TTT CTG ACT TGG TGT TTT CTC CTG AAT GCC TCC TTC 671672 CTG GCC CCC TAA TCA GAA ATG CAG AAG CCA CCT GCC CCA CAC AGG GGG 7191 Met Gln Lys Pro Pro Ala Pro His Arg Gly 10720 CCG CTT TCC CTG CAT CTG GCA CTC ACC AGG ACA GTC ATA GAA CTG GCC 76711 Pro Leu Ser Leu His Leu Ala Leu Thr Arg Thr Val Ile Glu Leu Ala 26768 TTG GGT GCT TGG GGA GAG TGG AGA GGT CTG GGG GGC ATC CAC TGG CCT 81527 Leu Gly Ala Trp Gly Glu Trp Arg Gly Leu Gly Gly Ile His Trp Pro 42816 CAG CCT CAG AGC TTC TCA TCA GTA ACC CAG GAG GCA AAG GCC CAG CTC 86343 Gln Pro Gln Ser Phe Ser Ser Val Thr Gln Glu Ala Lys Ala Gln Leu 58864 GAG ATT AGC CCA GGC CAC TGT ATC TGT GCC AAG ACC TAC AGC TGT CCC 91159 Glu Ile Ser Pro Gly His Cys Ile Cys Ala Lys Thr Tyr Ser Cys Pro 74912 TGC AGG GCG TGC ATC TCC TGC AGC CCA GGC TAT CAC AAT GGT GTC CCT 95975 Cys Arg Ala Cys Ile Ser Cys Ser Pro Gly Tyr His Asn Gly Val Pro 90960 TTT GGT GGG GAT GTG AGG CCT GCT GCC TGC CTG GGA AGA AGC TGA CAG 100791 Phe Gly Gly Asp Val Arg Pro Ala Ala Cys Leu Gly Arg Ser *** 1051008 GAT TGT TGG TGC TGT GGG GAG AAG CTG GTG TAA CTG AGA CCC ATG AGG 10551056 GAG GCC CTT TGT GGG TCT GTG GCT GGG AGG AGC TCT GGG GAA GAT GGG 11031104 CCT TGA GAA ATC CCA GCT TAG CAC C CAG TTG CTC CAC TGA GTC 11511152 ACC CTA GAC TGC AAT CTC ACA GAT GTT CTA ACC ACC CTC TCC CTG GGG 11991200 AGC CCT TGG GGG TGC TGC AAG CCC AGT CCT CAC TCA CTG GCT TTG TGC 12471248 TTC TCT TCC CGA CTT CCT TTT TTT G CCT GTT GAG AAA TGT GGG TCC 12951296 AGG GCT GCG TGC TCC AGT GTG CGG AAG GAA GGG CAG CGT GCT CTT TGG 13431344 ACT GAG CTG GCT CAA GGG AAG CTG GGT CTG AGG GGA GAC TGA CCT CGA 13911392 CCC TTC TTA CTT AAT GAA CCC TGC GTC TCA GTC ACC AGT TAC 14391440 CTG GCC TGA TCA CTG TGT GCC ACA CCC CAT CAC ATC CAT CCC CAC AAG 14871488 AAT GCC AGG AGG AGG GGT CAT TGG CCT TTT TGC CAG CAC AAA CCT CTG 15351536 GAG GCC AGC TCC CAA GAC CCA GGC ACA CCT GCC AGA GTG CCT TAA 15831584 CTT GGG ATG TCT GCA AAC CTC TGT CCA TGG CCG AAC CCA CCC CAC TGC 16311632 CTG TTA CTG TAA ATA AAG GTC TAT TAG CAT GCC ACC ATG CCC CTC CTC 16791680 CGA TGC CC ATA CAG T GCT CAG CAC CTG GTG CAA AGT AAG CAC AAT 17271728 CTG GCT TCC AGC CAT CAC TGC TGT TGC TAC TAT CAC AAA TAT TAG GCC 17751776 CCT TCT AGA ATG CAA AGA TAA CCC AGG GCT GCT CGT TCT GTG TCC AGA 18231824 TCA CAG AGT TTT T TTT TAT CCT TAT CAA CTT TAT TGA GGG ATA ATT 18711872 TAC AGG CAA TAA ATC ATA TCC ATT TAA AAA AAA AAA AAA AAA AA 19152. PP4377蛋白A：核苷酸序列(SEQ ID NO:4)长度：2132bpGGACGAGACT TGATGGAGCC ACCTATACAG TGCTGTTCAT TGGCACAGGA GACGGCTGGC 60TGCTCAAGGC TGTGAGCCTG GGGCCCTGGG TTCACCTGAT TGAGGAGCTG CAGCTGTTTG 120ACCAGGAGCC CATGAGAAGC CTGGTGCTAT CTCAGAGCAA GAAGCTGCTC TTTGCCGGCT 180CCCGCTCTCA GCTGGTGCAG CTGCCCGTGG CCGACTGCAT GAAGTATCGC TCCTGTGCAG 240ACTGTGTCCT CGCCCGGGAC CCCTATTGCG CCTGGAGCGT CAACACCAGC CGCTGTGTGC 300CATGGGTGGC CACTCTGGAT CTCTACTGAT CCAGCATGTG ATGACCTCGG ACACTTCAGG 360CATCTGCAAC CTCCGTGGCA GTAAGAAAGT CAGGCCCACT CCCAAAAACA TCACGGTGGT 420GGCGGGCACA GACCTGGTGC TGCCCTGCCA CCTCTCCTCC AACTTGGCCC ATGCCCGCTG 480GACCTTTGGG GGCCGGGACC TGCCTGCGGA ACAGCCCGGG TCCTTCCTCT ACGATGCCCG 540GCTCCAGGCC TGGTTGTGAT GGCTGCCCAG CCCCGCCATG CCGGGGCCTA CCACTGCTTT 600TCAGAGGAGC AGGGGGCGCG GCTGGCTGCT GAAGGCTACC TTGTGGCTGT CGTGGCAGGC 660CCTGTCGGTG ACCTTGGAGG CCCGGCCCCC CCTGGAAAAC CTGGGGCTGG TGTGGCTGGC 720GGTGGTGGCC CTGGGGGCTG TGTGCCTGGT GCTGCTGCTG CTGGTGCTGT CATTGCGCCG 780GCGGCTGCGG GAAGAGCTGG AGAAAGGGGC CAAGCTACTG AGAGGACCTT GGTGTACCCC 840CTGGAGCTGC CCAAGGAGCC CACCAGTCCC CCCTTCCGCC CTGTCCTGAA CCAGATGAGA 900AACTTTGGGA TCCTGTCGTT ACTACTATTC AGATGGCTCC CTTAAGATAG TACCTGGGCA 960TGCCCGGTGC CAGCCCGGTG GGGGGCCCCC TTCGCCACCT CCAGGCATCC CAGGCCAGCC 1020TCTGCCTTCT CCAACTCGGC TTCACCTGGG GGGTGGGCGG AACTCAAATG CCAATGGTTA 1080CGTGCGCTTA CAACTAGGAG GGGAGGACCG GGGAGGGCTC GGGCACCCCC TGCCTGAGCT 1140CGCGGATGAA CTGAGACGCA AACTGCAGCA ACGCCAGCCA CTGCCCGACT CCAACCCCGA 1200GGAGTCATCA GTATGAGGGG AACCCCCACC GCGTCGGCGG GAAGCGTGGG AGGTGTAGCT 1260CCTACTTTTG CACAGGCACC AGCTACCTCA GGGACATGGC ACGGGCACCT GCTCTGTCTG 1320GGACAGATAC TGCCCAGCAC CCACCCGGCC ATGAGGACCT GCTCTGCTCA GCACGGGCAC 1380TGACCTTGGT GTGGCTCACC AGGGCACCAG CCTCGCAGAA GGCATCTTCC TCCTCTCTGT 1440GAATCACAGA CACGCGGGAC CCCAGCCGCC AAAACTTTTC AAGGCAGAAG TTTCAAGATG 1500TGTGTTTGTC TGTATTTGCA CATGTGTTTG TGTGTGTGTG TATGTGTGTG TCCACGCGCG 1560TGCGCGCTTG TGGCATAGCC TTCCTGTTTC TGTCAAGTCT TCCCTTGGCC TGGGTCCTCC 1620TGGTGAGTCA TTGGAGCTAT GAAGGGGAAG GGGTCGTATC ACTTTGTCTC TCCTACCCCC 1680ACTGCCCCGA GTGTCGGGCA GCGATGTACA TATGGAGGTG GGGTGGACAG GGTGCTGTGC 1740CCCTTCAGAG GGAGTGCAGG GCTTGGGGTG GGCCTAGTCC TGCTCCTAGG GCTGTGAATG 1800TTTTCAGGGT GGGGGGAGGG AGATGGAGCC TCCTGTGTGT TTGGGGGGAA GGGTGGGTGG 1860GGCCTCCCAC TTGGCCCCGG GGTTCAGTGG TATTTTATAC TTGCCTTCTT CCTGTACAGG 1920GCTGGGAAAA GCTGTGTGAA GGGAGAGAAG GGAAAGGGTG GGCCTGCTGT GGACAATGGC 1980ATACTCTCTT CCAGCCCTAG GAGGAGGGCT CCTAACAGTG TAACTTATTG TGTCCCCGCG 2040TATTTATTTG TTGTAAATAT TTGAGTATTT TTATATTGCC AAATAAAATG GGAAAAAAAA 2100AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA AA 2132B：氨基酸序列(SEQ ID NO:5)长度：152个氨基酸1 MCVSTRVRAC GIAFLFLSSL PLAWVLLVSH WSYEGEGVVS LCLSYPHCPE CRAAMYIWRW61 GGQGAVPLQR ECRAWGGPSP APRAVNVFRV GGGRWSLLCV WGEGWVGPPT WPRGSVVFYT121 CLLPVQGWEK LCEGREGKGW ACCGQWHTLF QPC. Nucleotide and amino acid combination sequence (SEQ ID NO: 6) Clone number: PP4377 Start codon: 1542 ATG Stop codon: 2000 TAG Protein molecular weight: 169141 GG ACG AGA CTT GAT GGA GCC ACC TAT ACA GTG CTG TTC ATT GGC ACA 4748 GGA GAC GGC TGG CTG CTC AAG GCT GTG AGC CTG GGG CCC TGG GTT CAC 9596 CTG ATT GAG GAG CTG CAG CTG TTT GAC CAG GAG CCC ATG AGA AGC CTG 143144 GTG CTA TCT CAG AGC AAG AAG CTG CTC TTT GCC GGC TCC CGC TCT CAG 191192 CTG GTG CAG CTG CCC GTG GCC GAC TGC ATG AAG TAT CGC TCC TGT GCA 239240 GAC TGT GTC CTC GCC CGG GAC CCC TAT TGC GCC TGG AGC GTC AAC ACC 287288 AGC CGC TGT GTG CCA TGG GTG GCC ACT CTG GAT CTC TAC TGA TCC AGC 335336 ATG TGA TGA CCT CGG ACA CTT CAG GCA TCT GCA ACC TCC GTG GCA GTA 383384 AGA AAG TCA GGC CCA CTC CCA AAA ACA TCA CGG TGG TGG CGG GCA CAG 431432 ACC TGG TGC TGC CCT GCC ACC TCT CCT CCA ACT TGG CCC ATG CCC GCT 479480 GGA CCT TTG GGG GCC GGG ACC TGC CTG CGG AAC AGC CCG GGT CCT TCC 527528 TCT ACG ATG CCC GGC TCC AGG CCT GGT TGT GAT GGC TGC CCA GCC CCG 575576 CCA TGC CGG GGC CTA CCA CTG CTT TTC AGA G GCA GGG GGC GCG GCT 623624 GGC TGC TGA AGG CTA CCT TGT GGC TGT CGT GGC AGG CCC TGT CGG TGA 671672 CCT TGG AGG CCC GGC CCC CCC TGG AAA ACC TGG GGC TGG TGT GGC TGG 719720 CGG TGG TGG CCC TGG GGG TCCG CTG T TGC TGC TGC TGC TGG TGC 767768 TGT CAT TGC GCC GGC GGC TGC GGG AAG AGC TGG AGA AAG GGG CCA AGC 815816 TAC TGA GAG GAC CTT GGT GTA CCC CCT GGA GCT GCC CAA GGA GCC CAC 863864 CAG TCC CCC CTT CCG CCC TGT CCT GAA CCA GAT GAG AAA CTT TGG GAT 911912 CCT GTC GTT ACT ACT ATT CAG ATG GCT CCC TTA AGA TAG TAC CTG GGC 959960 ATG CCC GGT GCC AGC CCG GTG GGG GGC CCC CTT CGC CAC CTC CAG GCA 10071008 TCC CAG GCC AGC CTC TGC CTT CTC CAA CTC GGC TTC ACC TGG GGG GTG 10551056 GGC GGA ACT CAA ATG CCA ATG GTT ACG TGC GCT TAC AAC TAG GAG GGG 11031104 AGG ACC GGG GAG GGC TCG GGC ACC CCC TGC CTG AGC TCG CGG ATG AAC 11511152 AAC TGC AGC A G GCC AGC CAC TGC CCG ACT CCA ACC CCG 11991200 AGG AGT CAT CAG TAT GAG GGG AAC CCC CAC CGC GTC GGC GGG AAG CGT 12471248 GGG AGG TGT AGC TCC TAC TTT TGC ACA GGC ACC AGC TAC CTC AGG GAC 12951296 ATG CCT GCA CGG G CTG TCT GGG ACA GAT ACT GCC CAG CAC CCA 13431344 CCC GGC CAT GAG GAC CTG CTC TGC TCA GCA CGG GCA CTG ACC TTG GTG 13911392 TGG CTC ACC AGG GCA CCA GCC TCG CAG AAG GCA TCT TCC TCC TCT CTG 14391440 GACGA ATC CGG GAC CCC AGC CGC CAA AAC TTT TCA AGG CAG 14871488 AAG TTT CAA GAT GTG TGT TTG TCT GTA TTT GCA CAT GTG TTT GTG TGT 15351536 GTG TGT ATG TGT GTG TCC ACG CGC GTG CGC GCT TGT GGC ATA GCC TTC 158Val S Thr Arg Val Arg Ala Cys Gly Ile Ala Phe 141584 CTG TTT CTG TCA AGT CTT CCC TTG GCC TGG GTC CTC CTG GTG AGT CAT 163115 Leu Phe Leu Ser Ser Ser Leu Pro Leu Ala Trp Val Leu Leu Val Ser His 301632 TGG AGC TAT GAA GGG GAA GGG GTC GTA TCA CTT TGT CTC TCC TAC CCC 167931 Trp Ser Tyr Glu Gly Glu Gly Val Ser Leu Cys Leu Ser Tyr Pro 461680 CAC TGC CCC GAG TGT CGG GCA GCG ATG TAC ATA TGG AGG TGG GGT GGA 172747 His Cys Pro Glu Cys Arg Ala Ala Met Tyr Ile Trp Arg Trp Gly Gly 621728 CAG GGT GCT GTG CCC CTT CAG AGG GAG TGC AGG GCT TGG GGT GGG CCT 177563 Gln Gly Ala Val Pro Leu Gln Arg Glu Cys Arg Ala Trp Gly Gly Pro 781776 AGT CCT GCT CCT AGG GCT GTG AAT GTT TTC AGG GTG GGG GGA GGG AGA 182379 Ser Pro Ala Pro Arg Ala Val Asn Val Phe Arg Val Gly Gly Gly Arg 941824 TGG AGC CTC CTG TGT GTT TGG GGG GAA GGG TGG GTG GGG CCT CCC ACT 187195 Trp Leu Leu Cys Val Trp Gly Glu Gly Trp Val Gly Pro Thr 1101872 TGG CCC CGG GGT TCA GTG GTA TTT TAT ACT TGC CTT CTT CCT GTA CAG 1919111 Trp Pro Arg Gly Ser Val Val Phe Tyr Thr Cys Leu Leu Pro Val Gln 1261920 GGC TGG GAA AAG CTG TGT GAA GGG AGA GAA GGG AAA GGG TGG GCC TGC 1967127 Gly Trp Glu Lys Leu Cys Glu Gly Arg Glu Gly Lys Gly Trp Ala Cys 1421968 TGT GGA CAA TGG CAT ACT CTC TTC CAG CCC TAG GAG GAG GGC TCC TAA3 201514 Cys Gly Gln Trp His Thr Leu Phe Gln Pro *** 1532016 CAG TGT AAC TTA TTG TGT CCC CGC GTA TTT ATT TGT TGT AAA TAT TTG 20632064 AGT ATT TTT ATA TTG CCA AAT AAA ATG GGA AAA AAA AAA AAA AAA AAA 21112112 AAA AAA AAA AAA AAA AAA AAA 21323． PP4522蛋白A：核苷酸序列(SEQ ID NO:7)长度：2035bpGGCACCTCGA GTTTTTTTTT TTTTTATTCC CAGGACATCA AGGAGACTTT CAATAGGTGT 60GTTCCCAGTC TTTGCCCTTC GTGACCCAGT GGCATCTGGT TCCCTGTCCC TGCTTCTCTT 120CGGTATCCCT CTCCTCTCCT TCCTTCCCCA GGACCTGAGT TTCCATCTCC TGGACCCTCC 180TCTCCTTCCC CTCAGCTTTT GCTTTTCCCT CTGGGAATAT CGTGGTCCCA CCCCCTGCCC 240GGTCCCCTTC CTCCAGGTGT GAAGAGGTAC AGCTGCAGCC CCCAGAGGTC TGGTCCCCTG 300ACCCGTGCCA ACCCCATAGC CATGACTTCC TGACAGATGC CATCGTGAGG AAAATGAGCC 360GGATGTTCTG TCAGGCTGCG AGAGTGGACC TGACGCTGGA CCCTGACACG GCTCACCCGG 420CCCTGATGCT GTCCCCTGAC CGCCGGGGGG TCCGCCTGGC AGAGCGGCGG CAGGAGGTTG 480CTGACCATCC CAAGCGCTTC TCGGCCGACT GCTGCGTACT GGGGGCCCAG GGCTTCCGCT 540CCGGCCGGCA CTACTGGGAG GTAGAGGTGG GCGGGCGGCG GGGCTGGGCG GTGGGTGCTG 600CCCGTGAATC AACCCATCAT AAGGAAAAGG TGGGCCCTGG GGGTTCCTCC GTGGGCAGCG 660GGGATGCCAG CTCCTCGCGC CATCACCATC GCCGCCGCCG GCTCCACCTG CCCCAGCAGC 720CCCTGCTCCA GCGGGAAGTG TGGTGCGTGG GCACCAACGG CAAACGCTAT CAGGCCCAGA 780GCTCCACAGA ACAGACGCTG CTGAGCCCCA GTGAGAAACC AAGGCGCTTT GGTGTGTACC 840TGGACTATGA AGCTGGGCGC CTGGGCTTCT ACAACGCAGA GACTCTAGCC CACGTGCACA 900CCTTCTCGGC TGCCTTCCTG GGCGAGCGTG TCTTTCCTTT CTTCCGGGTG CTCTCCAAGG 960GCACCCGCAT CAAGCTCTGC CCTTGATTAT CCTGCCACCC GCAGGGGCCC CTCTGTCAGC 1020ACTTGGGGGG TGGGTGGTGG AGGGTGGCCC GTAAGTTTGA GGGCTCAAAG GCTCTTCCCA 1080CTGCTTGTTA CTGTGTTGCT TCCCACTCCC CCTTGACCCC AGGCCCCTGC TTCTCCCTCT 1140AGGAGCCTAA AGAACCCTCC TGGCCTCCAG CTCAGCCTTC TCTCACCTAC TATGTCTGTC 1200CAACAGGTCT GCATGGGTCC CTGATAATGA GAACAGCTGC CTGGTCTTCT CTCCCAGTCT 1260GCCTAGCCCA GCCCTGGGAC TGGAATTTGA GTAGGGGATG AGGGGAAATT GTAATTTCAT 1320TCCTTAACTT CCTTTTCCCC ACCCCTGCTC TTCAACCTCT TTATCAGTTC TGAGGCTGGA 1380GGGTTTGGGC AAGGCAACAT CCCCATTCCA ATTCCATTTT CTGATGCAGA TTTTAGCTGA 1440GGGATTTGGA AGCCATTTGG GGAGGCAGGC TGGGCCAAAG GGTAGAGCTG GGTAATAAAT 1500GTCTATTCTC CTGGGGAGGA GGGATTCTAA ACTTTCCTTC CGTCCTCAAT TTCTACCTCC 1560ATAGACCGGC CAGAATTTAG CTTCACTTGA GAGAGATCTG GAATGGTCGC CATGATTGAA 1620ACCACGCACC ATTACATCAT CATTACATTA ATTACATCAA CATAAATTAT TTCTTCCCCC 1680TTCCCTTTTC CAGCACTCAA CCAAGGAGCA AAGCTCATCC CACCCCACAC CCCTCCCAGG 1740TCTGCTCACT GCCAGGCTCC TCTCCCCTTT GTTCAGTGGA GCTGGCTTTT CTCCCAGCCC 1800CTTTCCATGC CTTTCACTCC ATTTGGCAAG CTCTGAGGGG GAGCCTGGGG ACGGGTTTGG 1860GTCCCCAGGA GGAGAGCCTT GGGTATAATC TATTTTTCTA GGAGCCTCTT GCCTTGTCAC 1920TTGCAGCTTT CGCCCTCTGC TTTGATGGCT GAGGTGAACT CATGTTCTTT GGGAAAAGGG 1980AAGGCGTGCT GTGGAAATAA AATGTTTATT TGCTTCTAAA AAAAAAAAAA AAAAA 2035B：氨基酸序列(SEQ ID NO:8)长度：210个氨基酸1 MSRMFCQAAR VDLTLDPDTA HPALMLSPDR RGVRLAERRQ EVADHPKRFS ADCCVLGAQG61 FRSGRHYWEV EVGGRRGWAV GAARESTHHK EKVGPGGSSV GSGDASSSRH HHRRRRLHLP121 QQPLLQREVW CVGTNGKRYQ AQSSTEQTLL SPSEKPRRFG VYLDYEAGRL GFYNAETLAH181 VHTFSAAFLG ERVFPFFRVL SKGTRIKLCPC. Nucleotide and amino acid combination sequence (SEQ ID NO:9) Clone number: PP4522 Start codon: 354 ATG Stop codon: 986 TGA Protein molecular weight: 236071 GG CAC CTC GAG TTT TTT TTT TTT TTA TTC CCA GGA CAT CAA GGA GAC 4748 TTT CAA TAG GTG TGT TCC CAG TCT TTG CCC TTC GTG ACC CAG TGG CAT 9596 CTG GTT CCC TGT CCC TGC TTC TCT TCG GTA TCC CTC TCC TCT CCT TCC 143144 TTC CCC AGG ACC TGA GTT TCC ATC TCC TGG ACC CTC CTC TCC TTC CCC 191192 TCA GCT TTT GCT TTT CCC TCT GGG AAT ATC GTG GTC CCA CCC CCT GCC 239240 CGG TCC CCT TCC TCC AGG TGT GAA GAG GTA CAG CTG CAG CCC CCA GAG 287288 GTC TGG TCC CCT GAC CCG TGC CAA CCC CAT AGC CAT GAC TTC CTG ACA 335336 GAT GCC ATC GTG AGG AAA ATG AGC CGG ATG TTC TGT CAG GCT GCG AGA 3831 Met Ser Arg Met Phe Cys Gln Ala Ala Arg 10384 GTG GAC CTG ACC CTG GAC CCT GAC ACG GCT CAC CCG GCC CTG Val1 ATG 13 CTG 4 Asp Leu Thr Leu Asp Pro Asp Thr Ala His Pro Ala Leu Met Leu 26432 TCC CCT GAC CGC CGG GGG GTC CGC CTG GCA GAG CGG CGG CAG GAG GTT 47927 Ser Pro Asp Arg Arg Gly Val Arg Leu Ala Glu Arg Arg Gln Glu Val 42480 GCT GAC CAT CCC AAG CGC TTC TCG GCC GAC TGC TGC GTA CTG GGG GCC 52743 Ala Asp His Pro Lys Arg Phe Ser Ala Asp Cys Cys Val Leu Gly Ala 58528 CAG GGC TTC CGC TCC GGC CGG CAC TAC TGG GAG GTA GAG GTG GGC GGG 57559 Gln Gly Phe Arg Ser Gly Arg His Tyr Trp Glu Val Glu Val Gly Gly 74576 CGG CGG GGC TGG GCG GTG GGT GCT GCC CGT GAA TCA ACC CAT CAT AAG 62375 Arg Arg Gly Trp Ala Val Gly Ala Ala Arg Glu Ser Thr His His Lys 90624 GAA AAG GTG GGC CCT GGG GGT TCC TCC GTG GGC AGC GGG GAT GCC AGC 67191 Glu Lys Val Gly Pro Gly Gly Ser Ser Ser Val Gly Ser Gly Asp Ala Ser 106672 TCC TCG CGC CAT CAC CAT CGC CGC CGC CGG CTC CAC CTG CCC CAG CAG 719107 Ser Ser Arg His His His Arg Arg Arg Arg Leu His Leu Pro Gln Gln 122720 CCC CTG CTC CAG CGG GAA GTG TGG TGC GTG GGC ACC AAC GGC AAA CGC 767123 Pro Leu Leu Gln Arg Glu Val Trp Cys Val Gly Thr Asn Gly Lys Arg 138768 TAT CAG GCC CAG AGC TCC ACA GAA CAG ACG CTG CTG AGC CCC AGT GAG 815139 Tyr Gln Ala Gln Ser Ser Thr Glu Gln Thr Leu Leu Ser Pro Ser Glu 154816 AAA CCA AGG CGC TTT GGT GTG TAC CTG GAC TAT GAA GCT GGG CGC CTG 863155 Lys Pro Arg Arg Phe Gly Val Tyr Leu Asp Tyr Glu Ala Gly Arg Leu 170864 GGC TTC TAC AAC GCA GAG ACT CTA GCC CAC GTG CAC ACC TTC TCG GCT 911171 Gly Phe Tyr Asn Ala Glu Thr Leu Ala His Val His Thr Phe Ser Ala 186912 GCC TTC CTG GGC GAG CGT GTC TTT CCT TTC TTC CGG GTG CTC TCC AAG 959187 Ala Phe Leu Gly Glu Arg Val Phe Pro Phe Phe Arg Val Leu Ser Lys 202960 GGC ACC CGC ATC AAG CTC TGC CCT TGA TTA TCC TGC CAC CCG CAG GGG 1007203 Gly Thr Arg Ile Lys Leu Cys Pro *** 2111008 CCC CTC TGT CAG CAC TTG GGG GGT GGG TGG TGG AGG GTG GCC CGT AAG 10551056 TTT GAG GGC TCA AAG GCT CTT CCC ACT GCT TGT TAC TGT GTT GCT TCC 11031104 CAC TCC CCC TTG ACC CCA GGC CCC TGC TTC TCC CTC TAG GAG CCT AAA 11511152 GAA CCC TCC TGG CCT CCA GCT CAG CCT TCT CTC ACC TAC TAT GTC TGT 11991200 CCA ACA GGT CTG CAT GGG TCC CTG ATA ATG AGA ACA G GCC TGG TCT 12471248 TCT CTC CCA GTC TGC CTA GCC CAG CCC TGG GAC TGG AAT TTG AGT AGG 12951296 GGA TGA GGG GAA ATT GTA ATT TCA TTC CTT AAC TTC CTT TTC CCC ACC 13431344 CCT GCT CTT CAA CCT CTT TAT CAG TTC TGA GGC TG AGG GTT TGG GCA 13911392 AGG CAA CAT CCC CAT TCC AAT TCC ATT TTC TGA TGC AGA TTT TAG CTG 14391440 AGG GAT TTG GAA GCC ATT TGG GGA GGC AGG CTG GGC CAA AGG GTA GAG 14871488 CTG GGT AAT AAA TGT CTA G TGA TCC TCC T GGG ATT CTA AAC TTT 15351536 CCT TCC GTC CTC AAT TTC TAC CTC CAT AGA CCG GCC AGA ATT TAG CTT 15831584 CAC TTG AGA GAG ATC TGG AAT GGT CGC CAT GAT TGA AAC CAC GCA CCA 16311632 TTA CAT CAT CAT CAT TAC ATT AAT TAC ATC AAC ATA AAT TAT TTC TTC CCC 16791680 CTT CCC TTT TCC AGC ACT CAA CCA AGG AGC AAA GCT CAT CCC ACC CCA 17271728 CAC CCC TCC CAG GTC TGC TCA CTG CCA GGC TCC TCT CCC CTT TGT TCA 17751776 GTG GAG CTG GCT TTT CTC CCA GCC CCT TTC CAT GCC TTT CAC TCC ATT 18231824 TGG CAA GCT CTG AGG GGG AGC CTG GGG ACG GGT TTG GGT CCC CAG GAG 18711872 GAG AGC CTT GGG TAT AAT CTA TTT TTC TAG GAG CCT CTT GCC TTG TCA 19191920 CTT GCA TTC TGC T GCC TT GAT GGC TGA GGT GAA CTC ATG TTC 19671968 TTT GGG AAA AGG GAA GGC GTG CTG TGG AAA TAA AAT GTT TAT TTG CTT 20152016 CTA AAA AAA AAA AAA AAA AAA AA 20354. PP4583蛋白A：核苷酸序列(SEQ ID NO:10)长度：2142bpCCGAAACGGG GAAGTTGTCT TGTGTGGAGA GGTTAGTAGA GCAGCGCGCG CGTCACCAGA 60GTCGTTTCTC TTCGGAGTCT TAGGTGATCG AGGGTGTGCC CAGGGGGCGG GACTTGTTTG 120CGCCTCCCGT TCCCTCCCAA TTTCCAAACG TGTCACCCCG GCGCCGACGG CCCTGTGCAG 180GGGAAGCAGA TGGAGTTCAA GCTGGAGGCT CATCGCATCG TCAGCATCTC TCTGGGCAAG 240ATCTACAACT CGCGGGTCCA GCGCGGCGGC ATCAAGCTGC ATAAGAACCT CCTGGTCTCG 300CTGGTGCTGC GCAGCGCCCG CCAAGTCTAC CTGAGCGACC CGTGCCCCGG CCTCTACCTG 360GCCGGTCCCG CTGGGACCCC GGCGCCGCCA CCGCAGCAGC AGCCCGGGGA GCCGGCGGCC 420GGGCCACCCG CCGGCTGGGG AGAGCCGCCC CCGCCCGCCG CTCATGCCTC TTGGCCGGAG 480ACCGAGCCGC AGCCGGAGCG CTCCTCCGTC TCAGACGCGC CGCGGGTAGG GGACGAGGTG 540CCGGTGGCCA CGGTGACTGG AGTCGGGGAC GTTTTTCAGG GCGGAGAGGC GGACGCGACG 600GAAGCTGCCT GGAGCCGCGT GGAGGGGCCG CGCCAGGCGG CGGCCAGAGA AGCCGAGGGT 660ACCGCCGGAG GCTGGGGCGT CTTCCCCGAG GTATCTCGTG CCGCGCGCCG CCCCTGCGGC 720TGCCCCCTAG GCGGGGAGGA CCCGCCGGGT ACACCGGCCG CGACCCCCGG CGCTGCCTGC 780TGCTGCGCGC CGCGACCAGC GGAGGACGAG CCCCCCGCGC CGCCCGCGGT GTGCCCCAGG 840AAGCGCTGCG CGGCGGGGGT GGGCGGCGGC CCAGCGGGCT GCCCGGCGCC CGGCTCGACC 900CCGCTCAAGA AGCCCCGCCG GAACTTAGAG CAGCCGCCGA GTGGAGGAGA GGACGACGAC 960GCGGAGGAGA TGGAGACCGG GAACGTGGCT AACCTCATCA GCATCTTCGG TTCCAGTTTC 1020TCGGGACTCC TACGGAAAAG CCCCGGGGGC GGCAGAGAGG AAGAGGAGGG AGAGGAGAGC 1080GGTCCGGAAG CCGCCGAGCC CGGGCAGATC TGCTGCGATA AGCCGGTGCT GAGAGACATG 1140AACCCCTGGA GCACAGCCAT CGTGGCCTTC TGAGCCCTTG GCCCCCCTGC GGGGAGGAGG 1200TGGAGCAGCG GGCGTCCCCG AAGTGAGGGC CAGGCCCTTC CCGGCTGCGA GGACGCCCAG 1260AGACCGCGGG CGCTGAGCGC GTTGGGCAGA CTGGTTGCCT CTGGGCATCG CAAACTGCCC 1320CCGGGCGCAT CTGGCTTATC TGGAGACCTG GGAGCTTGAG GCTCATTGGA AGAGGACGAT 1380CGTTAACTTT TGTGTTTGTG TATGTCTGTG TATGTAAGTT TGTCTTGTGG CATTAAGACT 1440ATTTTGCTCT TCTGGAATGC ACCACTCCTT CCCCAGGGTT TAAGAGATTG GGGGCAGACA 1500GGGACTTTCC TCTGCCGGCT GCGTGGAGAA GGAAGCGGCT AAAAGGTTTG GGCCGGGGAG 1560TTCCCCATTC GTTCTCCGGG GAGGGAGGGA CTTTACACCT ACCCCTCACC GGAAAGCTAG 1620ACCCGCTTCA GGGCCAGGAG TGGCGTTTCC GCACAGGATT TCCTAAGACG AGAGGGATTT 1680AGCCAAGAGC ACAGACTTGG ATTCCTTCTG TCCCTCCCCA CCTTTCTCTC CCAATGAAGA 1740AACGTTTATA TCCTGTGATT ACTCTAGGAA AGGGGCGTAT TAACGGTCCT TTGCTTCCCC 1800GCAGGGGAGA AAAAGCTTGA CGAACTTCAC AGAAGAGTTC AAGCTTGGAA ATAATGGAGT 1860TTATAGAGAA AAGATATATT TTTAAAAGCT CTAGGTCAGA AGTACTACAC AGTGCTTTTA 1920AAAAGTGTTT AATGAGAGTT TACAGACAGA AGCCCGAAGT GGAAAGACCT TATGGTTTTG 1980TAGATATGGT GACTCCAGTC TTTGTTGTAT AAAGGTTGGG GGAGCTGATA AGGTTTTTGT 2040ACAGTATTTT CTCCTTCGTT GTATTGATTT TTGTATAAAA ATGTAACTTC TACGTGTCTA 2100ACACGTATTA AATATTTTGA AGCAAAAAAA AAAAAAAAAA AA 2142B：氨基酸序列(SEQ ID NO:11)长度：327个氨基酸1 MEFKLEAHRI VSISLGKIYN SRVQRGGIKL HKNLLVSLVL RSARQVYLSD PCPGLYLAGP61 AGTPAPPPQQ QPGEPAAGPP AGWGEPPPPA AHASWPETEP QPERSSVSDA PRVGDEVPVA121 TVTGVGDVFQ GGEADATEAA WSRVEGPRQA AAREAEGTAG GWGVFPEVSR AARRPCGCPL181 GGEDPPGTPA ATPGAACCCA PRPAEDEPPA PPAVCPRKRC AAGVGGGPAG CPAPGSTPLK241 KPRRNLEQPP SGGEDDDAEE METGNVANLI SIFGSSFSGL LRKSPGGGRE EEEGEESGPE301 AAEPGQICCD KPVLRDMNPW STAIVAFC． Nucleotide and amino acid combination sequence (SEQ ID NO: 12) Clone number: PP4583 Start codon: 190 ATG Stop codon: 1173 TGA Protein molecular weight: 336121 CCG AAA CGG GGA AGT TGT CTT GTG TGG AGA GGT TAG TAG AGC AGC GCG 4849 CGC GTC ACC AGA GTC GTT TCT CTT CGG AGT CTT AGG TGA TCG AGG GTG 9697 TGC CCA GGG GGC GGG ACT TGT TTG CGC CTC CCG TTC CCT CCC AAT TTC 144145 CAA ACG TGT CAC CCC GGC GCC GAC GGC CCT GTG CAG GGGA AAG CAG ATG 1921 Met 1193 GAG TTC AAG CTG GAG GCT CAT CGC ATC GTC AGC ATC TCT CTG GGC AAG 2402 Glu Phe Lys Leu Glu Ala His Arg Ile Val Ser Ile Ser Leu Gly Lys 17241 ATC TAC AAC TCG CGG GTC CAG CGC GGC GGC ATC AAG CTG CAT AAG AAC 28818 Ile Tyr Asn Ser Arg Val Gln Arg Gly Gly Ile Lys Leu His Lys Asn 33289 CTC CTG GTC TCG CTG GTG CTG CGC AGC GCC CGC CAA GTC TAC CTG AGC 33634 Leu Leu Val Ser Leu Val Leu Arg Ser Ala Arg Gln Val Tyr Leu Ser 49337 GAC CCG TGC CCC GGC CTC TAC CTG GCC GGT CCC GCT GGG ACC CCG GCG 38450 Asp Pro Cys Pro Gly Leu Tyr Leu Ala Gly Pro Ala Gly Thr Pro Ala 65385 CCG CCA CCG CAG CAG CAG CCC GGG GAG CCG GCG GCC GGG CCA CCC GCC 43266 Pro Pro Pro Gln Gln Gln Gln Pro Gly Glu Pro Ala Ala Gly Pro Pro Ala 81433 GGC TGG GGA GAG CCG CCC CCG CCC GCC GCT CAT GCC TCT TGG CCG GAG 48082 Gly Trp Gly Glu Pro Pro Pro Pro Pro Ala Ala His Ala Ser Trp Pro Glu 97481 ACC GAG CCG CAG CCG GAG CGC TCC TCC GTC TCA GAC GCG CCG CGG GTA 52898 Thr Glu Pro Gln Pro Glu Arg Ser Ser Val Ser Asp Ala Pro Arg Val 113529 GGG GAC GAG GTG CCG GTG GCC ACG GTG ACT GGA GTC GGG GAC GTT TTT 576114 Gly Asp Glu Val Pro Val Ala Thr Val Thr Gly Val Gly Asp Val Phe 129577 CAG GGC GGA GAG GCG GAC GCG ACG GAA GCT GCC TGG AGC CGC GTG GAG 624130 Gln Gly Gly Glu Ala Asp Ala Thr Glu Ala Ala Trp Ser Arg Val Glu 145625 GGG CCG CGC CAG GCG GCG GCC AGA GAA GCC GAG GGT ACC GCC GGA GGC 672146 Gly Pro Arg Gln Ala Ala Ala Arg Glu Ala Glu Gly Thr Ala Gly Gly 161 673 TGG GGC GTC TTC CCC GAG GTA TCT CGT GCC GCG CGC CGC CCC TGC GGC 720162 Trp Gly Val Phe Pro Glu Val Ser Arg Ala Ala Arg Arg Pro Cys Gly 177721 TGC CCC CTA GGC GGG GAG GAC CCG CCG GGT ACA CCG GCC GCG ACC CCC 768178 Cys Pro Leu Gly Gly Glu Asp Pro Pro Gly Thr Pro Ala Ala Thr Pro 193769 GGC GCT GCC TGC TGC TGC GCG CCG CGA CCA GCG GAG GAC GAG CCC CCC 816194 Gly Ala Ala Cys Cys Cys Cys Ala Pro Arg Pro Ala Glu Asp Glu Pro Pro 209817 GCG CCG CCC GCG GTG TGC CCC AGG AAG CGC TGC GCG GCG GGG GTG GGC 864210 Ala Pro Pro Ala Val Cys Pro Arg Lys Arg Cys Ala Ala Gly Val Gly 225865 GGC GGC CCA GCG GGC TGC CCG GCG CCC GGC TCG ACC CCG CTC AAG AAG 912226 Gly Gly Pro Ala Gly Cys Pro Ala Pro Gly Ser Thr Pro Leu Lys Lys 241913 CCC CGC CGG AAC TTA GAG CAG CCG CCG AGT GGA GGA GAG GAC GAC GAC 960242 Pro Arg Arg Asn Leu Glu Gln Pro Pro Ser Gly Gly Glu Asp Asp Asp 257961 GCG GAG GAG ATG GAG ACC GGG AAC GTG GCT AAC CTC ATC AGC ATC TTC 1008258 Ala Glu Glu Met Glu Thr Gly Asn Val Ala Asn Leu lle Ser Ile Phe 2731009 GGT TCC AGT TTC TCG GGA CTC CTA CGG AAA AGC CCC GGG GGC GGC AGA 1056274 Gly Ser Ser Phe Ser Gly Leu Leu Arg Lys Ser Pro Gly Gly Gly Arg 2891057 GAG GAA GAG GAG GGA GAG GAG AGC GGT CCG GAA GCC GCC GAG CCC GGG 1104290 Glu Glu Glu Glu Gly Glu Glu Ser Gly Pro Glu Ala Ala Glu Pro Gly 3051105 CAG ATC TGC TGC GAT AAG CCG GTG CTG AGA GAC ATG AAC CCC TGG AGC 1152306 Gln Ile Cys Cys Asp Lys Pro Val Leu Arg Asp Met Asn Pro Trp Ser 3211153 ACA GCC ATC GTG GCC TTC TGA GCC CTT GGC CCC CCT GCG GG AGG AGG 1200322 Thr Ala Ile Val Ala Phe *** 3281201 TGG AGC AGC GGG CGT CCC CGA AGT GAG GGC CAG GCC CTT CCC GGC TGC 12481249 GAG GAC GCC CAG AGA CCG CGG GCG CTG AGC GCG TTG GGC AGA CTG TG TTT 12961 G97 CAT CGC AAA CTG CCC CCG GGC GCA TCT GGC TTA TCT GGA 13441345 GAC CTG GGA GCT TGA GGC TCA TTG GAA GAG GAC GAT CGT TAA CTT TTG 13921393 TGT TTG TGT ATG TCT GTG TAT GTA AGT TTG TCT TGT GGC ATT AAG ACT 14401TTG41 A CTC TTC TGG AAT GCA CCA CTC CTT CCC CAG GGT TTA AGA GAT 14881489 TGG GGG CAG ACA GGG ACT TTC CTC TGC CGG CTG CGT GGA GAA GGA AGC 15361537 GGC TAA AAG GTT TGG GCC GGG GAG TTC CCC ATT CGT TCT CCG GGAG 1 AGG 15 G84 GGA CTT TAC ACC TAC CCC TCA CCG GAA AGC TAG ACC CGC TTC AGG 16321633 GCC AGG AGT GGC GTT TCC GCA CAG GAT TTC CTA AGA CGA GAG GGA TTT 16801681 AGC CAA GAG CAC AGA CTT GGA TTC CTT CTG TCC CTC CCC ACC TTT CTC 17281729 TCC CAA TGA AGA AAC GTT TAT ATC CTG TGA TTA CTC TAG GAA AGG GGC 17761777 GTA TTA ACG GTC CTT TGC TTC CCC GCA GGG GAG AAA AAG CTT GAC GAA 18241825 CTT CAC AGA AGA AGA GTT CAA GCT TGG AAA TAA TGG AGT TTA TAG AGA AAA 18721873 GAT ATA TTT TTA AAA GCT CTA GGT CAG AAG TAC TAC ACA GTG CTT TTA 19201921 AAA AGT GTT TAA TGA GAG TTT ACA GAC AGA AGC CCG AAG TGG AAA GAC 19681969 CTT ATG GTT TTG TAG ATA TGG TGA CTC CAG TCT TTG AA TTG T GGT 20162017 TGG GGG AGC TGA TAA GGT TTT TGT ACA GTA TTT TCT CCT TCG TTG TAT 20642065 TGA TTT TTG TAT AAA AAT GTA ACT TCT ACG TGT CTA ACA CGT ATT AAA 21122113 TAT TTT GAA GCA AAA AAA AAA AAA AAA AAA 21425. PP4664蛋白A：核苷酸序列(SEQ ID NO:13)长度：2158bpGCCATCCCTA AGCAGACCCC AGTCCAATAC CTGCTCCCTG AGGCCAAGGC CCAGGACTCA 60GACAAGATCT GCGTGGTCAT CGACCTGGAC GAGACCCTGG TGCACAGCTC CTTCAAGCCA 120GTGAACAACG CGGACTTCAT CATCCCTGTG GAGATTGATG GGGTGGTCCA CCAGGTCTAC 180GTGTTGAAGC GTCCTCATGT GGATGAGTTC CTGCAGCGAA TGGGCGAGCT CTTTGAATGT 240GTGCTGTTCA CTGCTAGCCT CGCCAAGTAC GCAGACCCAG TAGCTGACCT GCTGGACAAA 300TGGGGGGCCT TCCGGGCCCG GCTGTTTCGA GAGTCCTGCG TCTTCCACCG GGGGAACTAC 360GTGAAGGACC TGAGCCGGTT GGGTCGAGAC CTGCGGCGGG TGCTCATCCT GGACAATTCA 420CCTGCCTCCT ATGTCTTCCA TCCAGACAAT GCTGTACCGG TGGCCTCGTG GTTTGACAAC 480ATGAGTGACA CAGAGCTCCA CGACCTCCTC CCCTTCTTCG AGCAACTCAG CCGTGTGGAC 540GACGTGTACT CAGTGCTCAG GCAGCCACGG CCAGGGAGCT AGTGAGGGTG ATGGGGCCAG 600GACCTGCCCC TGACCAATGA TACCCACACC TCCTCCCAGG AAGACTGCCC AGGCCTTTGT 660TAGGAAAACC CATGGGCCGC CGCCACACTC AGTGCCATGG GGAAGCGGGC GTCTCCCCCA 720CCAGCCCCAC CAGGCGGTGT AGGGGCAGCA GGCTGCACTG AGGACCGTGA GCTCCAGGCC 780CCGTGTCAGT GCCTTCAAAC CTCCTCCCCT ATTCTCAGGG GACCTGGGGG GCCCTGCCTG 840CTGCTCCCTT TTTCTGTCTC TGTCCATGCT GCCATGTTTC TCTGCTGCCA AATTGGGCCC 900CTTGGCCCCT TCCGGTTCTG CTTCCTGGGG GCAGGGTTCC TGCCTTGGAC CCCCAGTCTG 960GGAACGGTGG ACATCAAGTG CCTTGCATAG AGCCCCCTCT TCCCCGCCCA GCTTTCCCAG 1020GGGCACAGCT CTAGGCTGGG AGGGGAGAAC CAGCCCCTCC CCCTGCCCCA CCTCCTCCCT 1080TGGGACTGAG AGGGCCCCTA CCAACCTTTG CCTCTGCCTT GGAGGGAGGG GAGGTCTGTT 1140ACCACTGGGG AAGGCAGCAG GAGTCTGTCC TTCAGGCCCC ACAGTGCAGC TTCTCCAGGG 1200CCGACAGCTG AGGGCTGCTC CCTGCATCAT CCAAGCAATG ACCTCAGACT TCTGCCTTAA 1260CCAGCCCCGG GGCTTGGCTC CCCCAGCTCT GAGCGTGGGG GCATAGGCAG GACCCCCCTT 1320GTGGTGCCAT ATAAATATGT ACATGTGTAT ATAGATTTTT AGGGGAAGGA GAGAGGGAAG 1380GGTCAGGGTA GAGACACCCC TCCCTTGCCC CTTTCCTGGG CCCAGAAGTT GGGGGGAGGG 1440AGGGAAAGGA TTTTTACATT TTTTAAACTG CTATTTTCTG AATGGAACAA GCTGGGCCAA 1500GGGGCCCAGG CCCTGTCCTC TGTCCCTCAC ACCCCTTTGC TCCGTTCATT CATTCAAAAA 1560AACATTTCTT GAGCACCTTC TGTGCCCAGC ATATGCTAGG CCCACCAGCT AAGTGTGTGT 1620GGGGGGTCTC TACGCCAGCT CATCAGTGCC TCCTTGCCCA TCCTTCACCG GTGCCTTTGG 1680GGGATCTGTA GGAGGTGGGA CCTTCTGTGG GGTTTGGGGA TCTCCAGGAA GCCCGACCAA 1740GCTGTCCCCT TCCCCTGTGC CAACCCATCT CCTACAGCCC CCTGCCTGAT CCCCTGCTGG 1800CTGGGGGCAG CTCCCAGGAT ATCCTGCCTT CCAACTGTTT CTGAAGCCCC TCCTCCTAAC 1860ATGGCGATTC CGGAGGTCAA GGCCTTGGGC TCTCCCCAGG GTCTAACGGT TAAGGGGACC 1920CACATACCAG TGCCAAGGGG GATGTCAAGT GGTGATGTCG TTGTGCTCCC CTCCCCCAGA 1980GCGGGTGGGC GGGGGGTGAA TATGGTTGGC CTGCATCAGG TGGCCTTCCC ATTTAAGTGC 2040CTTCTCTGTG ACTGAGAGCC CTAGTGTGAT GAGAACTAAA GAGAAAGCCA GACCCCTAAA 2100AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA AAAAAAAA 2158B：氨基酸序列(SEQ ID NO:14)长度：146个氨基酸1 MLPCFSAAKL GPLAPSGSAS WGQGSCLGPP VWERWTSSAL HRAPSSPPSF PRGTALGWEG61 RTSPSPCPTS SLGTERAPTN LCLCLGGRGG LLPLGKAAGV CPSGPTVQLL QGRQLRAAPC121 IIQAMTSDFC LNQPRGLAPP ALSVGAC. Nucleotide and amino acid combination sequence (SEQ ID NO: 15) Clone number: PP4664 Start codon: 866 ATG Stop codon: 1306 TAG Protein molecular weight: 148151 G CCA TCC CTA AGC AGA CCC CAG TCC AAT ACC TGC TCC CTG AGG CCA 4647 AGG CCC AGG ACT CAG ACA AGA TCT GCG TGG TCA TCG ACC TGG ACG AGA 9495 CCC TGG TGC ACA GCT CCT TCA AGC CAG TGA ACA ACG CGG ACT TCA TCA 142 143 TCC CTG TGG AGA TTG ATG GGG TGG TCC ACC GAC AGG TCT TGT TGA AGC 190191 GTC CTC ATG TGG ATG AGT TCC TGC AGC GAA TGG GCG AGC TCT TTG AAT 238239 GTG TGC TGT TCA CTG CTA GCC TCG CCA AGT ACG CAG ACC CAG TAG CTG 286287 ACC TGC TGG ACA AAT GGG GGG CCT TGC GGCC TGT TTC GAG AGT 334335 CCT GCG TCT TCC ACC GGG GGA ACT ACG TGA AGG ACC TGA GCC GGT TGG 382383 GTC GAG ACC TGC GGC GGG TGC TCA TCC TGG ACA ATT CAC CTG CCT CCT 430431 ATG TCT TCC ATC CAGA ACA ATG CTG TAC CGG TG CCT CGT GGT TTG ACA 478479 ACA TGA GTG ACA CAG AGC TCC ACG ACC TCC TCC CCT TCT TCG AGC AAC 526527 TCA GCC GTG TGG ACG ACG TGT ACT CAG TGC TCA GGC AGC CAC GGC CAG 574575 GGA GCT AGT GAG GGT GAT GGG GCC AGG ACC TGC CCC TGA CCA ATG ATA 622623 CCC ACA CCT CCT CCC AGG AAG ACT GCC CAG GCC TTT GTT AGG AAA ACC 670671 CAT GGG CCG CCG CCA CAC TCA GTG CCA TGG GGA AGC GGG CGT CTC CCC 718719 CAC CAG CCC CAC CAG GCG GTG TAG GG CAG CAG GCT GCA CTG AGG ACC 766767 GTG AGC TCC AGG CCC CGT GTC AGT GCC TTC AAA CCT CCT CCC CTA TTC 814815 TCA GGG GAC CTG GGG GGC CCT GCC TGC TGC TCC CTT TTT CTG TCT CTG 862863 TCC ATG CTG CCA TGT TTC TCT GCT GCC AAA TTG GGC CCC TTG GCC CCT 9101 Met Leu Pro Cys Phe Ser Ala Ala Lys Leu Gly Pro Leu Ala Pro 15911 TCC GGT TCT GCT TCC TGG GGG CAG GGT TCC TGC CTT GGA CCC CCA GTC 95816 Ser Gly Ser Ala Ser Trp Gly Gln Gly Ser Cys Leu Gly Pro Pro Val 31959 TGG GAA CGG TGG ACA TCA AGT GCC TTG CAT AGA GCC CCC TCT TCC CCG 100632 Trp Glu Arg Trp Thr Ser Ser Ser Ala Leu His Arg Ala Pro Ser Ser Pro 471007 CCC AGC TTT CCC AGG GGC ACA GCT CTA GGC TGG GAG GGG AGA ACC AGC 105448 Pro Ser Phe Pro Arg Gly Thr Ala Leu Gly Trp Glu Gly Arg Thr Ser 631055 CCC TCC CCC TGC CCC ACC TCC TCC TCC CTT GGG ACT GAG AGG GCC CCT ACC 110264 Pro Ser Pro Cys Pro Thr Ser Ser Leu Gly Thr Glu Arg Ala Pro Thr 791103 AAC CTT TGC CTC TGC CTT GGA GGG AGG GGA GGT CTG TTA CCA CTG GGG 115080 Asn Leu Cys Leu Cys Leu Gly Gly Arg Gly Gly Leu Pro Leu Gly 951151 AAG GCA GCA GGA GTC TGT CCT TCA GGC CCC ACA GTG CAG CTT CTC CAG 119896 Lys Ala Ala Gly Val Cys Pro Ser Gly Pro Thr Val Gln Leu Leu Gln 1111199 GGC CGA CAG CTG AGG GCT GCT CCC TGC ATC ATC CAA GCA ATG ACC TCA 1246112 Gly Arg Gln Leu Arg Ala Ala Pro Cys Ile Ile Gln Ala Met Thr Ser 1271247 GAC TTC TGC CTT AAC CAG CCC CGG GGC TTG GCT CCC CCA GCT CTG AGC 1294128 Asp Phe Cys Leu Asn Gln Pro Arg Gly Leu Ala Pro Pro Ala Leu Ser 1431295 GTG GGG GCA TAG GCA GGA CCC CCC TTG TGG TGC CAT ATA AAT ATG TAC 1342144 Val Gly Ala *** 1471343 ATG TGT ATA TAG ATT TTT AGG GGA AGG AGA GAG GGA AGG GTC AGG GTA 13901391 GAG ACA CCC CTC CCT TGC CCC TTT CCT GGG CCC AGA AGT TGG GGG GAG 14381439 GGA GGG AAA GGA TTT TTA CAT TTT TTA AAC TGC TAT TTT CTG AAT GGA 14861487 ACA AGC TGG GCC AAG GGG CCC AGG CCC TGT CCT CTG TCC CTC ACA CCC 15341535 CTT TGC TCC GTT CAT TCA TTC AAA AAA A CTT GAG CAC CTT CTG 15821583 TGC CCA GCA TAT GCT AGG CCC ACC AGC TAA GTG TGT GTG GGG GGT CTC 16301631 TAC GCC AGC TCA TCA GTG CCT CCT TGC CCA TCC TTC ACC GGT GCC TTT 16781679 GGG GGA TCT GGTTA GGA GGT GGG ACC T GGG GTT TGG GGA TCT CCA 17261727 GGA AGC CCG ACC AAG CTG TCC CCT TCC CCT GTG CCA ACC CAT CTC CTA 17741775 CAG CCC CCT GCC TGA TCC CCT GCT GGC TGG GGG CAG CTC CCA GGA TAT 18221823 CCT GCC TTC CAA CTAG TTT CCG A CTC CTC CTA ACA TGG CGA TTC 18701871 CGG AGG TCA AGG CCT TGG GCT CTC CCC AGG GTC TAA CGG TTA AGG GGA 19181919 CCC ACA TAC CAG TGC CAA GGG GGA TGT CAA GTG GTG ATG TCG TTG TGC 19661967 TCC CCTGC CCC C GGG GGG TGA ATA TGG TTG GCC TGC 20142015 ATC AGG TGG CCT TCC CAT TTA AGT GCC TTC TCT GTG ACT GAG AGC CCT 20622063 AGT GTG ATG AGA ACT AAA GAG AAA GCC AGA CCC CTA AAA AAA AAA AAA 21102111 AAA AAA AAA AAA AAA A AAA AAA AAA AAA AAA AAA AAA AAA AAA 21586. PP4748蛋白A：核苷酸序列(SEQ ID NO:16)长度：1337bpGAACAGCGGG GGCACACGGG GCGCTGCCGA AGTGCAAGGC CACGGCCAGA GCTCGAGCCC 60GACGCGCTGT CTGGAGTCGT AGGTTGGCGC CGTTTGGGGT CGGGGTCTGA GGCTTGGGCG 120CTGCCTGGGC CGAGCGGAGA TCGGGGTTTG CCTCCCGTCC CCGCTCAGGA CCCTGACGTG 180GCTGAAGCGG CCCCGGGAGC ATGAGCGGGC AGCGCGTGGA CGTCAAGGTG GTGATGCTGG 240GCAAGGAGTA CGTGGGCAAG ACTAGCCTGG TGGAGCGCTA CGTGCACGAC CGCTTTCTGG 300TGGGGCCTTA TCAGAACACC ATCGGGGCCG CCTTCGTGGC CAAGGTGATG TCGGTCGGAG 360ACCGGACTGT GACATTAGGT ATTTGGGACA CAGCAGGCTC TGAGCGCTAT GAGGCCATGA 420GTAGAATCTA CTATCGGGGT GCCAAGGCTG CCATCGTCTG CTATGACCTC ACAGACAGCA 480GCAGCTTTGA GCGAGCAAAG TTCTGGGTGA AGGAACTGCG CAGCCTAGAG GAGGGCTGCC 540AAATCTACTT ATGTGGCACC AAGAGTGACC TGCTGGAAGA AGACCGGAGG CGTCGACGTG 600TGGACTTCCA CGACGTCCAG GACTATGCAG ACAGTAGCTG CTCCTCAGCC CTTTGGGGGG 660TGGGGGTGTG TGGCTGTCTG GGTGGATCAA AGAAAATAGG GACTGCCTTG GCTGCCAGGG 720CAAGGTGCTC TAGGAGGTCT TCCTGGCCTC CTTGAACTGT GGGGTCCAGG AGACTCCCTG 780AACTGCTAGC CCTCCCTTTT GTCTGTTTAT CTAATTCTCA GGTATGAGGC TTTAGTCACT 840TCTCTTTACA GATATCAAAG CTCAGCTCTT TGAAACATCC AGCAAGACAG GCCAGAGTGT 900GGACGAGCTC TTCCAGAAAG TGGCAGAGGA TTACGTCAGT GTGGCTGCCT TCCAGGTGAT 960GACAGAGGAC AAGGGCGTGG ATCTGGGCCA GAAGCCAAAC CCCTACTTCT ACAGCTGTTG 1020TCATCACTGA GTCAGCACTC ACCTGGCCTG GGGGAATTAA AGGAATTCCC CGTAAGGGCT 1080GGACCCAGCT CCTTTCTGGG CTTGGGTAGT CAAATGTCTG AGCTACCCCA GGTCCTCATG 1140TCAGCAGAGT GGCGCCTGCC TGTGCTGGCC CATGGAACGG AGACAGCATT GGGCTGACTG 1200TGGGCATGAG GAGGGATAAG GCTGATTTGG ACCCCAGGCT TCTGCCCTGG ACAGCACTTG 1260TGTCTGCAGA TTATTTAAGT GGCTTTTGAT CTGTAAATAA AATCAGTGCA CTGTGCATCA 1320AAAAAAAAAA AAAAAAA 1337B：氨基酸序列(SEQ ID NO:17)长度：184个氨基酸1 MSGQRVDVKV VMLGKEYVGK TSLVERYVHD RFLVGPYQNT IGAAFVAKVM SVGDRTVTLG61 IWDTAGSERY EAMSRIYYRG AKAAIVCYDL TDSSSFERAK FWVKELRSLE EGCQIYLCGT121 KSDLLEEDRR RRRVDFHDVQ DYADSSCSSA LWGVGVCGCL GGSKKIGTAL AARARCSRRS181 SWPPC． Nucleotide and amino acid combination sequence (SEQ ID NO: 18) Clone number: PP4748 Start codon: 201 ATG Stop codon: 755 TGA Protein molecular weight: 205171 GA ACA GCG GGG GCA CAC GGG GCG CTG CCG AAG TGC AAG GCC ACG GCC 4748 AGA GCT CGA GCC CGA CGC GCT GTC TGG AGT CGT AGG TTG GCG CCG TTT 9596 GGG GTC GGG GTC TGA GGC TTG GGC GCT GCC TGG GCC GAG CGG AGA TCG 143144 GGG TTT GCC TCC CGT CCC CGC TCA GGA CCC TGA CGT GGC TGA AGC GGC 191192 CCC GGG AGC ATG AGC GGG CAG CGC GTG GAC GTC AAG GTG GTG ATG CTG 2391 Met Ser Gly Gln Arg Val Asp Val Lys Val Val Met Leu 13 240 GGC AAG GAG TAC GTG GGC AAG ACT AGC CTG GTG GAG CGC TAC GTG CAC 28714 Gly Lys Glu Tyr Val Gly Lys Thr Ser Leu Val Glu Arg Tyr Val His 29288 GAC CGC TTT CTG GTG GGG CCT TAT CAG AAC ACC ATC GGG GCC GCC TTC 33530 Asp Arg Phe Leu Val Gly Pro Tyr Gln Asn Thr Ile Gly Ala Ala Phe 45336 GTG GCC AAG GTG ATG TCG GTC GGA GAC CGG ACT GTG ACA TTA GGT ATT 38346 Val Ala Lys Val Met Ser Val Gly Asp Arg Thr Val Thr Leu Gly Ile 61384 TGG GAC ACA GCA GGC TCT GAG CGC TAT GAG GCC ATG AGT AGA ATC TAC 43162 Trp Asp Thr Ala Gly Ser Glu Arg Tyr Glu Ala Met Ser Arg Ile Tyr 77432 TAT CGG GGT GCC AAG GCT GCC ATC GTC TGC TAT GAC CTC ACA GAC AGC 47978 Tyr Arg Gly Ala Lys Ala Ala Ile Val Cys Tyr Asp Leu Thr Asp Ser 93480 AGC AGC TTT GAG CGA GCA AAG TTC TGG GTG AAG GAA CTG CGC AGC CTA 52794 Ser Ser Phe Glu Arg Ala Lys Phe Trp Val Lys Glu Leu Arg Ser Leu 109528 GAG GAG GGC TGC CAA ATC TAC TTA TGT GGC ACC AAG AGT GAC CTG CTG 575110 Glu Glu Gly Cys Gln Ile Tyr Leu Cys Gly Thr Lys Ser Asp Leu Leu 125576 GAA GAA GAC CGG AGG CGT CGA CGT GTG GAC TTC CAC GAC GTC CAG GAC 623126 Glu Glu Asp Arg Arg Arg Arg Arg Val Asp Phe His Asp Val Gln Asp 141624 TAT GCA GAC AGT AGC TGC TCC TCA GCC CTT TGG GGG GTG GGG GTG TGT 671142 Tyr Ala Asp Ser Ser Ser Cys Ser Ser Ala Leu Trp Gly Val Gly Val Cys 157672 GGC TGT CTG GGT GGA TCA AAG AAA ATA GGG ACT GCC TTG GCT GCC AGG 719158 Gly Cys Leu Gly Gly Ser Lys Lys Ile Gly Thr Ala Leu Ala Ala Arg 173720 GCA AGG TGC TCT AGG AGG TCT TCC TGG CCT CCT TGA ACT GTG GGG TCC 767174 Ala Arg Cys Ser Arg Arg Ser Ser Trp Pro Pro *** 185768 AGG AGA CTC CCT GAA CTG CTA GCC CTC CCT TTT GTC TGT TTA TCT AAT 815816 TCT CAG GTA TGA GGC TTT AGT CAC TTC TCT TTA CAG ATA TCA AAG CTC 863864 AGC TCT TTG AAA CAT CCA GCA AGA CAG GCC AGA GTG TGG ACG AGC TCT 911912 TCC AGA AAG TGG CAG AGG ATT ACG TCA GTG TGG CTG CCT TCC AGG TGA 959960 TGA CAG AGG ACA AGG GCG TGG ATC TGG GCC AGA AGC CAA ACC CCT ACT 10071008 TCT ACA GTC GTT G GTC TCA GCA CTC ACC TGG CCT GGG GGA 10551056 ATT AAA GGA ATT CCC CGT AAG GGC TGG ACC CAG CTC CTT TCT GGG CTT 11031104 GGG TAG TCA AAT GTC TGA GCT ACC CCA GGT CCT CAT GTC AGC AGA GTG 11511152 GCG CCT GCC TCAGT GCT GGC C TGG AAC GGA GAC AGC ATT GGG CTG ACT 11991200 GTG GGC ATG AGG AGG GAT AAG GCT GAT TTG GAC CCC AGG CTT CTG CCC 12471248 TGG ACA GCA CTT GTG TCT GCA GAT TAT TTA AGT GGC TTT TGA TCT GTA 12951296 AGT G AAA A TC TGC ATC AAA AAA AAA AAA AAA AAA 1337D. Blastp result Query=PP4748 protein (184 amino acids)>SP_IN:P91857 P91857 caenorhabditis elegans.f26h9.6 protein.11/1999

Length=208 amino acid score=108bits(266), expected value=3e-23 identity=60/140(42%), similarity=88/140(62%), gap=8/140(5%) Query:9 KVVMLGKEYVGKTSLVERYVHDRFLVGPYQ-NTIGAAFVAKVMSVGDRTVTLGIWDTAGS 67

     K+V+LG+  VGK+SLV R+V  +F    YQ +TIGAAF+ + + + D T+   IWDTAGSbjct:21 KLVLLGESAVGKSSLVLRFVKGQF--HEYQESTIGAAFLTQTVCLDDATIKFEIWDTAGQ 78Query: 68  ERYEAMSRIYYRGAKAAIVCYDLTDSSSFERAKFWVKEL-RSLEEGCQIYLCGTKSXXXX 126

ERY +++ +YYRGA+AAIV YD+T+ SF++AK WVKEL R + L G K+Sbjct: 79 ERYHSLAPMYYRGAQAAIVVYDITNQESFQKAKNWVKELQRQASSPNIVMALAGNKA---- 134Query: 127 XXXADXXXX1

V++ + YA+ +Sbjct: 135 DVANKRTVEYEEANAYAEDN 1547． PP4762蛋白A：核苷酸序列(SEQ ID NO:19)长度：2417bpGTTTCTGGCG CGAACTTCCG CCGTTCCGAA GTTGCACGGT GAATTGGCGC TATGTCTGGG 60GACAGCAGCG GCCGCGGGCC AGAGGGCCGG GGCCGGGGCC GCGACCCGCA TCGGGATCGC 120ACCCGCTCCC GCTCCCGCTC GCGGTCCCCT TTGTCGCCCA GGTCCCGCCG CGGCTCTGCG 180CGGGAGCGCA GAGAGGCCCC AGAGCGCCCG AGCCTGGAGG ACACAGAGCC GTCGGATTCC 240GGGGACGAGA TGATGGACCC GGCCAGCTTG GAGGCGGAGG CCGACCAAGG CCTGTGCCGC 300CAGATCCGCC ATCAGTACCG GGCGCTCATC AACTCCGTCC AACGTAAGGC GGCGCCTCCG 360GGCGGCGCGG GCCCGGACGG GCCGCTGTCC CCGCCCTGCG CCGGGCGCGG GGGGAGGGCA 420CCGGAGGCAG GTGCCGGGCG GAGGCCCGGC GCGGCGTTAG TAGAGACGCG CCGGGTGCCA 480GCCCCTCGCC GCACCTTAAA TTGTTGACAA AGTGCTCCCA CTTTGTGGCT GGAGCCGTAC 540GACCTTGGCG CGGGTCCCCG AGTGGCGGGC TGGAAAAGCT GGAGGCCAGA GCCGTTCGCG 600GACTCGCCCG AGCCCAGCCA GCCAGTGGGG AGAGGCAGGA CGCGGACTCC GCGTGCTTTG 660ATTGCGCCCG GCAGTCCTCG GCGTGGGCTC CCGGCCTCGC AGCGGCACGC TGAGCTGCAG 720CGGCACGCGA GAGCTGTCAA ATGCACGTTC TCGGACCCCA CCCGGACCCG CGGAATTCGA 780ATCTCTGGGT GGGTCCGCGC AACTGTGGAG TGGCCCTCCA GGTGATGGTG TGTATTGCTT 840GCAAAGTTTG AGAACCACTC AGTTATGGAG CACTCAGAGG CTGCAGCCGG AGTCCAGCCC 900TTTGGGTTAA GGTTAGAGGG AGCTAGGGTT TCCTTTTAAA ATTCATGCTG CATAAAGGGT 960AGGGTGGCCT TTCTCCTTTT CCGGGTCATT TTGGTTAAAA AGATAAAGAT GTTCGAAGTA 1020ATTACTCAGA GCGCAGGGGC CGGGGCTTGG CTTTACTGTT GCAGGTGATA ACCTTGTCAA 1080CTGCTTATTA GTGAATGCTG AACAAATTTC CCAAAGTTTG TATTTTTAAA AATAGAAAAC 1140CGTGAGACAT ACTGAATGCC GGTGACAAAT TAACAGAGGT CCTTGAAGAG GCTAACACTC 1200TGTTTAATGA AGTGTCCCGA GCAAGAGAAG CAGTCCTGGA TGCCCACTTT CTTGTTTTGG 1260CTTCAGATTT GGGCAAAGAG AAAGCAAAGC AGCTGCGCTC AGACCTGAGC TCCTTTGACA 1320TGTTAAGATA TGTTGAAACT CTACTCACAC ATATGGGTGT AAATCCGCTA GAAGCTGAAG 1380AACTCATCCG TGATGAAGAT AGTCCTGATT TTGAATTCAT AGTCTATGAC TCCTGGAAGA 1440TAACAGGCAG AACAGCAGAA AACACCTTTA ATAAAACCCA TACATTCCAC TTTCTGTTGG 1500GTTCAATATA CGGAGAGTGC CCTGTGCCAA AGCCACGAGT TGATCGTCCA AGAAAAGTTC 1560CTGTGATACA AGAGGAGAGG GCAATGCCTG CCCAGTTAAG AAGAATGGAA GAATCTCATC 1620AAGAAGCAAC AGAGAAAGAA GTAGAAAGAA TCTTGGGATT GTTGCAGACA TATTTTCGAG 1680AAGATCGAGA CAGTCTCACT CTTTCGCCCA GACTGGAATG CAGTGGCACC ATCTCAACTC 1740ACTGCAACCT CCACCTCCTG GATTCAAGCA ATTCTCCTGC CTCAGCCTCC TGAGTAGCTG 1800GGATTACAGG CGCACGCGCC ACCACATCTG GCTAATTTTT TTTCTGTATT TTTAGTAGAG 1860ACATTGTTTC GCCATGTTGG CCAAGCTAGT CTTGAACCCC TGACCTCAGG TGATCTGCCA 1920GCATCGGCCT CCCAAAGTGC TGGGATTACA GGTCTGATAC CCCAATGTCC TTCTTTGACT 1980TTGTGGTTGA TCCTCATTCT TTCCCCCGTA CAGTGGAAAA CATCTTTCAT GTTTCCTTCA 2040TTATACGGGA TGGTTTTGCA AGAATAAGAC TTGACCAAGA CCGACTGCCA GTAATAGAGC 2100CTGTTAGTAT TAATGAAGAA AATGAGGGAT TTGAACATAA CACACAAGTT AGAAATCAAG 2160GAATTATAGC TTTGAGTTAC CGTGACTGGG AGGAGATTGT GAAGACCTTT GAGATTTCAG 2220AGCCTGTGAT TACTCCAAGT CAGAGGCAGC AGAAGCCAAG TGCTTGATGC TAGCTGAAGG 2280ACTCAAATGG ATAGTGAAGT CCAAAACGGA AAGCGGCATG TATTGTACAT ATTGTATGAT 2340TCAACATTTT TAAAGGCAGA TTGTTTTTAG TAAAATGTAG CTTTTGATAG TTAAAAAAAA 2400AAAAAAAAAA AAAAAAA 2417B：氨基酸序列(SEQ ID NO:20)长度：157个氨基酸1 MLRYVETLLT HMGVNPLEAE ELIRDEDSPD FEFIVYDSWK ITGRTAENTF NKTHTFHFLL61 GSIYGECPVP KPRVDRPRKV PVIQEERAMP AQLRRMEESH QEATEKEVER ILGLLQTYFR121 EDRDSLTLSP RLECSGTIST HCNLHLLDSS NSPASASC． Nucleotide and amino acid combination sequence (SEQ ID NO: 21) Clone number: PP4762 Start codon: 1320 ATG Stop codon: 1793 TGA Protein molecular weight: 180601 GT TTC TGG CGC GAA CTT CCG CCG TTC CGA AGT TGC ACG GTG AAT TGG 4748 CGC TAT GTC TGG GGA CAG CAG CGG CCG CGG GCC AGA GGG CCG GGG CCG 9596 GGG CCG CGA CCC GCA TCG GGA TCG CAC CCG CTC CCG CTC CCG CTC GCG 143144 GTC CCC TTT GTC GCC CAG GTC CCG CCG CGGCG GTCG GTCG T GCG CAG 191192 AGA GGC CCC AGA GCG CCC GAG CCT GGA GGA CAC AGA GCC GTC GGA TTC 239240 CGG GGA CGA GAT GAT GGA CCC GGC CAG CTT GGA GGC GGA GGC CGA CCA 287288 AGG CCT GTG CCG CCA GAT CCG CCA TCA GTA CCG GGC GCT CAT CAA CTC 335336 CGT CCA ACG TAA GGC GGC GCC TCC GGG CGG CGC GGG CCC GGA CGG GCC 383384 GCT GTC CCC GCC CTG CGC CGG GCG CGG GGG GAG GGC ACC GGA GGC AGG 431432 TGC CGG GCG GAG GCC CGG CGC GGC GACTT AGT AGA GCG CCG GGT GCC 479480 AGC CCC TCG CCG CAC CTT AAA TTG TTG ACA AAG TGC TCC CAC TTT GTG 527528 GCT GGA GCC GTA CGA CCT TGG CGC GGG TCC CCG AGT GGC GGG CTG GAA 575576 AAG CTG GAG GCC AGA GCC GTT CGC GGA GTC GTC CGA GCC CAG CCA GCC 623624 AGT GGG GAG AGG CAG GAC GCG GAC TCC GCG TGC TTT GAT TGC GCC CGG 671672 CAG TCC TCG GCG TGG GCT CCC GGC CTC GCA GCG GCA CGC TGA GCT GCA 719720 GCG GCA CGC GAGA AGC TGT CAA ATG CAC GTT CTC GGA CCC CAC CCG GAC 767768 CCG CGG AAT TCG AAT CTC TGG GTG GGT CCG CGC AAC TGT GGA GTG GCC 815816 GTC CAG GTG ATG GTG TGT ATT GCT TGC AAA GTT TGA GAA CCA CTC AGT 863864 TAT GGA GCA CTC AGA GGC TGC AGC CG AGT CCA GCC CTT TGG GTT AAG 911912 GTT AGA GGG AGC TAG GGT TTC CTT TTA AAA TTC ATG CTG CAT AAA GGG 959960 TAG GGT GGC CTT TCT CCT TTT CCG GGT CAT TTT GGT TAA AAA GAT AAA 10071008 GAT GTT CGA AGT AAT TAC TCA GAG CGC AGG GGC CGG GGC TTG GCT TTA 10551056 CTG TTG CAG GTG ATA ACC TTG TCA ACT GCT TAT TAG TGA ATG CTG AAC 11031104 AAA TTT CCC AAA GTT TGT ATT TTT AAA AAT AGA AAA CCG TGA GAC ATA 11511152 CTGA AACAT A GAA T GTA G ACA GAG GTC CTT GAA GAG GCT AAC ACT 11991200 CTG TTT AAT GAA GTG TCC CGA GCA AGA GAA GCA GTC CTG GAT GCC CAC 12471248 TTT CTT GTT TTG GCT TCA GAT TTG GGC AAA GAG AAA GCA AAG CAG CTG 12951296 CGC TCA GAC CTG AGC T TTT GAC ATG TTA AGA TAT GTT GAA ACT CTA 13431 Met Leu Arg Tyr Val Glu Thr Leu 81344 CTC ACA CAT ATG GGT GTA AAT CCG CTA GAA GCT GAA GAA CTC ATC CGT 13919 Leu Thr His Met Gly Val Asn Pro Leu Glu Ala Glu Glu Leu Ile Arg 241392 GAT GAA GAT AGT CCT GAT TTT GAA TTC ATA GTC TAT GAC TCC TGG AAG 143925 Asp Glu Asp Ser Pro Asp Phe Glu Phe Ile Val Tyr Asp Ser Trp Lys 401440 ATA ACA GGC AGA ACA GCA GAA AAC ACC TTT AAT AAA ACC CAT ACA TTC 148741 Ile Thr Gly Arg Thr Ala Glu Asn Thr Phe Asn Lys Thr His Thr Phe 561488 CAC TTT CTG TTG GGT TCA ATA TAC GGA GAG TGC CCT GTG CCA AAG CCA 153557 His Phe Leu Leu Gly Ser Ile Tyr Gly Glu Cys Pro Val Pro Lys Pro 721536 CGA GTT GAT CGT CCA AGA AAA GTT CCT GTG ATA CAA GAG GAG AGG GCA 158373 Arg Val Asp Arg Pro Arg Lys Val Pro Val Ile Gln Glu Glu Arg Ala 881584 ATG CCT GCC CAG TTA AGA AGA ATG GAA GAA TCT CAT CAA GAA GCA ACA 163189 Met Pro Ala Gln Leu Arg Arg Met Glu Glu Ser His Gln Glu Ala Thr 1041632 GAG AAA GAA GTA GAA AGA ATC TTG GGA TTG TTG CAG ACA TAT TTT CGA 1679105 Glu Lys Glu Val Glu Arg Ile Leu Gly Leu Leu Gln Thr Tyr Phe Arg 1201680 GAA GAT CGA GAC AGT CTC ACT CTT TCG CCC AGA CTG GAA TGC AGT GGC 1727121 Glu Asp Arg Asp Ser Leu Thr Leu Ser Pro Arg Leu Glu Cys Ser Gly 1361728 ACC ATC TCA ACT CAC TGC AAC CTC CAC CTC CTG GAT TCA AGC AAT TCT 1775137 Thr Ile Ser Thr His Cys Asn Leu His Leu Leu Asp Ser Ser Asn Ser 1521776 CCT GCC TCA GCC TCC TGA GTA GCT GGG ATT ACA GGC GCA CGC GCC ACC 1823153 Pro Ala Ser Ala Ser ** * 1581824 ACA TCT GGC TAA TTT TTT TTC TGT ATT TTT AGT AGA GAC ATT GTT TCG 18711872 CCA TGT TGG CCA AGC TAG TCT TGA ACC CCT GAC CTC AGG TGA TCT GCC 19191920 AGC ATC GGC CTC CCA AAG TGC TGG GAT TAC AGG TCT GAT ACC CCA ATG 19671968 TCC TTC TTT GAC TTT GTG GTT GAT CCT CAT TCT TTC CCC CGT ACA GTG 20152016 GAA AAC ATC TTT CAT GTT TCC TTC ATT ATA CGG GAT GGT TTT GCA AGA 20632064 ATA AGA CTT GAC CAA GAC CGA CTG CCA GTA ATA GAG CCT GTT AGT ATT 21112112 AAT GAA GAA AAT GAG GGA TTT GAA CAT AAC ACA CAA GTT AGA AAT CAA 21592160 GGA ATT ATA GCT TTG AGT TAC CGT GAC TGG GAG GAG ATT GTG AAG ACC 22072208 TTT GAG ATT TCA GAG CCT GTG ATT ACT CCA AGT CAG AGG CAG CAG AAG 22552256 CCA AGT GCT TGA TGC TAG CTG AAG GAC TCA AAT GGA TAG TGA AGT CCA 23032304 AAA CGG AAA GCG GCA TGT ATT GTA CAT ATT GTA TGA TTC AAC ATT TTT 23512352 AAA GGC AGA TTG TTT TTA GTA AAA TGT AGC TTT TGA TAG TTA AAA AAA 23992400 AAA AAA AAA AAA AAA AAA 24178. PP5242蛋白A：核苷酸序列(SEQ ID NO:22)长度：1685bpGCACCGCGGA GGACAGGGGC AGCTGGCGGG CAGCGGGTGA GGGGGTGGCG GGGACGCGAG 60TGGCGGCCGC GGGGCCCCGG ACAAGGGTCC GCAGAGCTGC AGCCTTCGAG GGCCAGCCCT 120CTCCGAGTCC GGGGCTGGGT CCCACCAGTG ACAAGGCGGC AGCCCCGCGC ACACCAAAGA 180GAAGGCGGCT GTGGCGGCAG CGGCAGCCCC AGCCATGCTG TGTTATGTGA CGAGGCCGGA 240CGCGGTGCTG ATGGAGGTGG AGGTGGAGGC GAAAGCCAAC GGCGAGGACT GCCTCAACCA 300GGTGTGCAGG CGACTGGGAA TCATAGAAGT TGACTATTTT GGACTGCAGT TTACGGGTAG 360CAAAGGTGAA AGTTTATGGC TAAACCTGAG AAACCGGATC TCCCAGCAGA TGGATGGGCT 420AGCCCCTTAC AGGCTTAAAC TTAGAGTCAA GTTCTTCGTG GAGCCTCATC TCATCTTACA 480GGAGCAGACT AGGCATATCT TTTTCTTGCA CATCAAGGAG GCCCTCTTGG CAGGCCACCT 540CTTGTGTTCC CCAGAGCAGG CAGTGGAACT CAGTGCCCTC CTGGCCCAGA CCAAGTTTGG 600AGACTACAAC CAGAACACTG CCAAGTATAA CTATGAGGAG CTCTGTGCCA AGGAGCTCTC 660CTCTGCCACC TTGAACAGCA TTGTTGCAAA ACATAAGGAG TTGGAGGGGA CCAGCCAGGC 720TTCAGCTGAA TACCAAGTTT TGCAGATTGT GTCGGCAATG GAAAACTATG GCATAGAATG 780GCATTCTGTG CGGGATAGCG AAGGGCAGAA ACTGCTCATT GGGGTTGGAC CTGAAGGAAT 840CTCAATTTGT AAAGATGACT TTAGCCCAAT TAATAGGATA GCTTATCCTG TGGTGCAGAT 900GGCCACCCAG TCAGGAAAGA ATGTATATTT GACGGTCACC AAGGAATCTG GGAACAGCAT 960CGTGCTCTTG TTTAAAATGA TCAGCACCAG GGCGGCCAGC GGGCTCTACC GAGCGATAAC 1020AGAGACGCAC GCATTCTACA GGTGTGACAC AGTGACCAGC GCCGTGATGA TGCAGTATAG 1080CCGTGACTTG AAGGGCCACT TGGCATCTCT GTTTCTGAAT GAAAACATTA ACCTTGGCAA 1140GAAATATGTC TTTGATATTA AAAGAACATC AAAGGAGGTG TATGACCATG CCAGGAGGGC 1200TCTGTACAAT GCTGGCGTTG TGGACCTCGT TTCAAGAAGC AACCAGAGCC CTTCACACTC 1260GCCTCTGAAG TCCTCAGAAA GCAGCATGAA CTGCAGCAGC TGCGAGGGCC TCAGCTGCCA 1320GCAGACCCGG GTGCTGCAGG AGAAGCTACG CAAGCTGAAG GAAGCCATGC TGTGCATGGT 1380GTGCTGCGAG GAGGAGATCA ACTCCACCTT CTGTCCCTGT GGCCACACTG TGTGCTGTGA 1440GAGCTGCGCC GCCCAGCTAC AGTCATGTCC CGTCTGCAGG TCGCGTGTGG AGCATGTCCA 1500GCACGTCTAT CTGCCAACGC ACACCAGTCT TCTCAATCTG ACTGTAATCT AATCTGTTGT 1560GCTTTTGTTG GACTTGGCAT GTTTCCATGA ACTGCACTAT TATAAACTAT TAAAATGATA 1620GATTGTGGAG AAAGTAATTA TTCCAACACC CATCTGCCAT GCGATGTTAA AAAAAAAAAA 1680AAAAA 1685B：氨基酸序列(SEQ ID NO:23)长度：445个氨基酸1 MLCYVTRPDA VLMEVEVEAK ANGEDCLNQV CRRLGIIEVD YFGLQFTGSK GESLWLNLRN61 RISQQMDGLA PYRLKLRVKF FVEPHLILQE QTRHIFFLHI KEALLAGHLL CSPEQAVELS121 ALLAQTKFGD YNQNTAKYNY EELCAKELSS ATLNSIVAKH KELEGTSQAS AEYQVLQIVS181 AMENYGIEWH SVRDSEGQKL LIGVGPEGIS ICKDDFSPIN RIAYPVVQMA TQSGKNVYLT241 VTKESGNSIV LLFKMISTRA ASGLYRAITE THAFYRCDTV TSAVMMQYSR DLKGHLASLF301 LNENINLGKK YVFDIKRTSK EVYDHARRAL YNAGVVDLVS RSNQSPSHSP LKSSESSMNC361 SSCEGLSCQQ TRVLQEKLRK LKEAMLCMVC CEEEINSTFC PCGHTVCCES CAAQLQSCPV421 CRSRVEHVQH VYLPTHTSLL NLTVIC． Nucleotide and amino acid combination sequence (SEQ ID NO: 24) Clone number: PP5242 Start codon: 215 ATG Stop codon: 1552 TAA Protein molecular weight: 498811 G CAC CGC GGA GGA CAG GGG CAG CTG GCG GGC AGC GGG TGA GGG GGT 4647 GGC GGG GAC GCG AGT GGC GGC CGC GGG GCC CCG GAC AAG GGT CCG CAG 9495 AGC TGC AGC CTT CGA GGG CCA GCC CTC TCC GAG TCC GGG GCT GGG TCC 142143 CAC CAG TGA CAA GGC GGC AGC CCC GCG CAC ACC AAA GAG AAG GCG GCT 190191 GTG GCG GCA GCG GCA GCC CCA GCC ATG CTG TGT TAT GTG ACG AGG CCG 2381 Met Leu Cys Tyr Val Thr Arg Pro 8239 GAC GCG GTG CTG ATG GAG GTG GAG GTG GAG GCG AAA GCC AAC GGC GAG 2869 Asp Ala Met Glu Val Glu Val Glu Ala Lys Ala Asn Gly Glu 24287 GAC TGC CTC AAC CAG GTG TGC AGG CGA CTG GGA ATC ATA GAA GTT GAC 33425 Asp Cys Leu Asn Gln Val Cys Arg Arg Leu Gly Ile Ile Glu Val Asp 40335 TAT TTT GGA CTG CAG TTT ACG GGT AGC AAA GGT GAA AGT TTA TGG CTA 38241 Tyr Phe Gly Leu Gln Phe Thr Gly Ser Lys Gly Glu Ser Leu Trp Leu 56383 AAC CTG AGA AAC CGG ATC TCC CAG CAG ATG GAT GGG CTA GCC CCT TAC 43057 Asn Leu Arg Asn Arg Ile Ser Gln Gln Met Asp Gly Leu Ala Pro Tyr 72431 AGG CTT AAA CTT AGA GTC AAG TTC TTC GTG GAG CCT CAT CTC ATC TTA 47873 Arg Leu Lys Leu Arg Val Lys Phe Phe Val Glu Pro His Leu Ile Leu 88479 CAG GAG CAG ACT AGG CAT ATC TTT TTC TTG CAC ATC AAG GAG GCC CTC 52689 Gln Glu Gln Thr Arg His Ile Phe Phe Leu His Ile Lys Glu Ala Leu 104527 TTG GCA GGC CAC CTC TTG TGT TCC CCA GAG CAG GCA GTG GAA CTC AGT 574105 Leu Ala Gly His Leu Leu Cys Ser Pro Glu Gln Ala Val Glu Leu Ser 120 575 GCC CTC CTG GCC CAG ACC AAG TTT GGA GAC TAC AAC CAG AAC ACT GCC 622121 Ala Leu Leu Ala Gln Thr Lys Phe Gly Asp Tyr Asn Gln Asn Thr Ala 136623 AAG TAT AAC TAT GAG GAG CTC TGT GCC AAG GAG CTC TCC TCT GCC ACC 670137 Lys Tyr Asn Tyr Glu Glu Leu Cys Ala Lys Glu Leu Ser Ser Ala Thr 152671 TTG AAC AGC ATT GTT GCA AAA CAT AAG GAG TTG GAG GGG ACC AGC CAG 718153 Leu Asn Ser Ile Val Ala Lys His Lys Glu Leu Glu Gly Thr Ser Gln 168719 GCT TCA GCT GAA TAC CAA GTT TTG CAG ATT GTG TCG GCA ATG GAA AAC 766169 Ala Ser Ala Glu Tyr Gln Val Leu Gln Ile Val Ser Ala Met Glu Asn 184767 TAT GGC ATA GAA TGG CAT TCT GTG CGG GAT AGC GAA GGG CAG AAA CTG 814185 Tyr Gly Ile Glu Trp His Ser Val Arg Asp Ser Glu Gly Gly Gln Lys Leu 200815 CTC ATT GGG GTT GGA CCT GAA GGA ATC TCA ATT TGT AAA GAT GAC TTT 862201 Leu Ile Gly Val Gly Pro Glu Gly Ile Ser Ile Cys Lys Asp Asp Phe 216863 AGC CCA ATT AAT AGG ATA GCT TAT CCT GTG GTG CAG ATG GCC ACC CAG 910217 Ser Pro Ile Asn Arg Ile Ala Tyr Pro Val Val Gln Met Ala Thr Gln 232911 TCA GGA AAG AAT GTA TAT TTG ACG GTC ACC AAG GAA TCT GGG AAC AGC 958233 Ser Gly Lys Asn Val Tyr Leu Thr Val Thr Lys Glu Ser Gly Asn Ser 248959 ATC GTG CTC TTG TTT AAA ATG ATC AGC ACC AGG GCG GCC AGC GGG CTC 1006249 Ile Val Leu Leu Phe Lys Met Ile Ser Thr Arg Ala Ala Ser Gly Leu 2641007 TAC CGA GCG ATA ACA GAG ACG CAC GCA TTC TAC AGG TGT GAC ACA GTG 1054265 Tyr Arg Ala Ile Thr Glu Thr His Ala Phe Tyr Arg Cys Asp Thr Val 2801055 ACC AGC GCC GTG ATG ATG CAG TAT AGC CGT GAC TTG AAG GGC CAC TTG 1102281 Thr Ser Ala Val Met Met Gln Tyr Ser Arg Asp Leu Lys Gly His Leu 2961103 GCA TCT CTG TTT ACG AAT GAA A ATT AAC CTT GGC AAG AAA TAT GTC 1150297 Ala Ser Leu Phe Leu Asn Glu Asn Ile Asn Leu Gly Lys Lys Tyr Val 3121151 TTT GAT ATT AAA AGA ACA TCA AAG GAG GTG TAT GAC CAT GCC AGG AGG 1198313 Phe Asp Ile Lys Arg Thr Ser Lys Glu Val Tyr Asp His Ala Arg Arg 3281199 GCT CTG TAC AAT GCT GGC GTT GTG GAC CTC GTT TCA AGA AGC AAC CAG 1246329 Ala Leu Tyr Asn Ala Gly Val Val Asp Leu Val Ser Arg Ser Asn Gln 3441247 AGC CCT TCA CAC TCG CCT CTG AAG TCC TCA GAA AGC AGC ATG AAC TGC 1294345 Ser Pro Ser His Ser Pro Leu Lys Ser Ser Glu Ser Ser Met Asn Cys 3601295 AGC AGC TGC GAG GGC CTC AGC TGC CAG CAG ACC CGG GTG CTG CAG GAG 1342361 Ser Ser Cys Glu Gly Leu Ser Cys Gln Gln Thr Arg Val Leu Gln Glu 3761343 AAG CTA CGC AAG CTG AAG GAA GCC ATG CTG TGC ATG GTG TGC TGC GAG 1390377 Lys Leu Arg Lys Leu Lys Glu Ala Met Leu Cys Met Val Cys Cys Glu 3921391 GAG GAG ATC AAC TCC ACC TTC TGT CCC TGT GGC CAC ACT GTG TGC TGT 1438393 Glu Glu Ile Asn Ser Thr Phe Cys Pro Cys Gly His Thr Val Cys Cys 4081439 GAG AGC TGC GCC GCC CAG CTA CAG TCA TGT CCC GTC TGC AGG TCG CGT 1486409 Glu Ser Cys Ala Ala Gln Leu Gln Ser Cys Pro Val Cys Arg Ser Arg 4241487 GTG GAG CAT GTC CAG CAC GTC TAT CTG CCA ACG CAC ACC AGT CTT CTC 1534425 Val Glu His Val Gln His Val Tyr Leu Pro Thr His Thr Ser Leu Leu 4401535 AAT CTG ACT GTA ATC TAA TCT GTT GTG CTT TTG TTG GAC TTG GCA TGT 1582441 Asn Leu Thr Val Ile *** 4461583 TTC CAT GAA CTG CAC TAT TAT AAA CTA TTA AAA TGA TAG ATT GTG GAG 16301631 AAA GTA ATT ATT CCA ACA CCC ATC TGC CAT GCG ATG TTA AAA AAA AAA 16781679 AAA AAA A 1685D. Blastp result Query=PP5242 protein (445 amino acids)>SP_IN:Q24440 Q24440 drosophila melanogaster(fruit fly).(cdnal)

  protein 4.1 homologue (coracle). 11/1999

Length=1698 amino acids score=125bits(312), expected value=4e-28 identity=82/284(28%), similarity=138/284(47%), gap=8/284(2%) Query: 2 LCYVTRPDAVLMEVEAKANGEDCLNQVCRRLGIIEVDYFGLQFTGSKGESLWLNLRNR 61

       L  VT  D  L++V ++ KA G D +N +C  L +IE DYFGL +        WL+LSbjct: 33  LARVTLLDGSLLDVSIDRKAIGRDVINSICAGLNLIEKDYFGLTYETPTDPRTWLDLEKP  92Query: 62  ISQQMDGLAPYRLKLRVKFF-VEPHLILQEQTRHIFFLHIKEALLAGHLLCSPEQAVELS 120

       +S +      + L   VKF+  EP  + ++ TR+   L ++  +L G L C+      LSbjct: 93  VS-KFFRTDTWPLTFAVKFYPPEPSQLKEDITRYHLCLQVRNDILEGRLPCTFVTHALLG 151Query: 121 ALLAQTKFGDYNQN---TAKYNYEELCAKELSSATLNSIVAKHKELEGTSQASAEYQVLQ 177

       + L Q++ GDY+     T  Y  +   A   ++   + ++  HK  +G S A AE   L+Sbjct: 152 SYLVQSEMGDYDAEEMPTRAYLKDFKIAPNQTAELEDKVMDLHKTHKGQSPAEAELHYLE 211Query: 178 IVSAMENYGIEWHSVRDSEGQKLLIGVGPEGISICKDDFSPINRIAYPVVQMATQSGKNV 237

           +  YG++ H  +DSEG  +++GV   G+ + +D    INR A+P +   +    +Sbjct: 212 NAKKLAMYGVDLHPAKDSEGVDIMLGVCASGLLVYRDKLR-INRFAWPKILKISYKRHHF 270Query: 238 YLTVTKESGNSI--VLLFKMISTRAASGLYRAITETHAFYRCDT 279

Y+ + + FK+ + RAA L+++ E H F+R TSbjct: 271 YIKIRPGEFEQYESTIGFKLANHRAAKKLWKSCVEHHTFFRLMT 3149. PP5656蛋白A：核苷酸序列(SEQ ID NO:25)长度：1373bpTTCCGCAATT TCTACCTGAC CTTCCTGGAG TACGATGGGA ACCTGCTGCG GAGAGAGCTC   60TTTGTGTGCC CCAGCCAGCC CCCACCTGGT GCTGAGCAGT TGCAGCAGGC CCTGGCACAA  120CTGGACGAGG AAGACCCCTG CTTTGAGTTC CGGCAGCAGC AGCTCACTGT GCACCGTGTG  180CATGTCACTT TCCTGCCCCA TGAACCGCCA CCCCCCCGGC CTCACGATGT CACCCTTGTG  240GCCCAGCTGT CCATGGACCG GCTGCAGATG TTGGAAGCCC TGTGCAGGCA CTGGCCTGGC  300CCCATGAGCC TGGCCTTGTA CCTGACAGAC GCAGAAGCTC AGCAGTTCCT GATTTCGGTC  360GAGGCCTCAC CAGTGCTTGC TGCCCGGCAG GACGTGGCCT ACCATGTGGT GTACCGTGAG  420GGCCCCTATA CCCCGTCAAC CAGCTTCGCA ACGTGGCCTT GGCCAGGCCC TCACGCCTTA  480CGTCTTCCTC AGTGACATTG ACTTCCTGCC TGCCTATTCT CTCTACGACT ACCTCAGGGC  540CTCCATTGAG CAGCTGGGGC TGGGCAGCCG GCGCAAGGCA GCACTGGTGG TGCCGGCATT  600CGAGACCCTG CGCTACCGCT TCAGCTTCCC CCATTCCAAG GTGGAGCTGT TGGCCTTGCT  660GGATGCGGGC ACTCTCTACA CCTTCAGGTA CCACGAGTGG CCCCGAGGCC ACGCACCCAC  720AGACTATGCC CGCTGGCGGG AGGCTCAGGC CCCGTACCGT GTGCAATGGG CGGCCAACTA  780TGAACCCTAC GTGGTGGTGC CACGAGACTG TCCCCGCTAT GATCCTCGCT TTGTGGGCTT  840CGGCTGGAAC AAAGTGGCCC ACATTGTGGA GCTGGATGCC CAGGAATATG AGCTCCTGGT  900GCTGCCCGAG GCCTTCACCA TCCATCTGCC CCACGCTCCA AGCCTGGACA TCTCCGCTTC   960CGCTCCAGCC CCACCTATCG TGACTGCCTC CAGGCCCTCA AGGACGAATT CCACCAGGAC  1020TTGTCCCGCC ACCATGGGGC TGCTGCCCTC AAATACCTCC CAGCCCTGCA GCAGCCCCAG  1080AGCCCTGCCC GAGGCTGAGG CTGGGCCGGC GCTGCCCCTC ATCTTAGCAT TGGGCAGACA  1140CCAGGGCAAC CTGCCCTCCG CCATCCCTGC TATTTAAATT ATTTAAGGTC TCTGGGAAGG  1200GCTGGGGCAG AGCATCTGTG GGGTGGGGTC TTCCCCTTGC TGCTATTGTA TGGCTGGGGA  1260CTGGTCTCTC TCTGCCCCAG CCAGTTTGGG GCTGGTTCCC CCATCTTGAA TTGTTTATCC  1320CTTTTTCATA ATTAAAGTTT TAAAACATAA AAAAAAAAAA AAAAAAAAAA AAA         1373B：氨基酸序列(SEQ ID NO:26)长度：257个氨基酸1 MWCTVRAPIP RQPASQRGLG QALTPYVFLS DIDFLPAYSL YDYLRASIEQ LGLGSRRKAA61 LVVPAFETLR YRFSFPHSKV ELLALLDAGT LYTFRYHEWP RGHAPTDYAR WREAQAPYRV121 QWAANYEPYV VVPRDCPRYD PRFVGFGWNK VAHIVELDAQ EYELLVLPEA FTIHLPHAPS181 LDISASAPAP PIVTASRPSR TNSTRTCPAT MGLLPSNTSQ PCSSPRALPE AEAGPALPLI241 LALGRHQGNL PSAIPAIC． Nucleotide and amino acid combination sequence (SEQ ID NO: 27) clone number: PP5656 Start codon: 404 ATG Termination codon: 1177 TAA Protein molecular weight: 285571 T TCC GCA ATT TCT ACC TGA CCT TCC TGG AGT ACG ATG GGA ACC TGC      4647  TGC GGA GAG AGC TCT TTG TGT GCC CCA GCC AGC CCC CAC CTG GTG CTG      9495  AGC AGT TGC AGC AGG CCC TGG CAC AAC TGG ACG AGG AAG ACC CCT GCT     142143  TTG AGT TCC GGC AGC AGC AGC TCA CTG TGC ACC GTG TGC ATG TCA CTT     190191  TCC TGC CCC ATG AAC CGC CAC CCC CCC GGC CTC ACG ATG TCA CCC TTG     238239  TGG CCC AGC TGT CCA TGG ACC GGC TGC AGA TGT TGG AAG CCC TGT GCA     286287  GGC ACT GGC CTG GCC CCA TGA GCC TGG CCT TGT ACC TGA CAG ACG CAG     334335  AAG CTC AGC AGT TCC TGA TTT CGG TCG AGG CCT CAC CAG TGC TTG CTG     382383  CCC GGC AGG ACG TGG CCT ACC ATG TGG TGT ACC GTG AGG GCC CCT ATA     4301                              Met Trp Cys Thr Val Arg Ala Pro Ile       9431  CCC CGT CAA CCA GCT TCG CAA CGT GGC CTT GGC CAG GCC CTC ACG CCT     47810  Pro Arg Gln Pro Ala Ser Gln Arg Gly Leu Gly Gln Ala Leu Thr Pro      25479  TAC GTC TTC CTC AGT GAC ATT GAC TTC CTG CCT GCC TAT TCT CTC TAC     52626  Tyr Val Phe Leu Ser Asp Ile Asp Phe Leu Pro Ala Tyr Ser Leu Tyr      41527  GAC TAC CTC AGG GCC TCC ATT GAG CAG CTG GGG CTG GGC AGC CGG CGC     57442  Asp Tyr Leu Arg Ala Ser Ile Glu Gln Leu Gly Leu Gly Ser Arg Arg      57575 AAG GCA GCA CTG GTG GTG CCG GCA TTC GAG ACC CTG CGC TAC CGC TTC     62258  Lys Ala Ala Leu Val Val Pro Ala Phe Glu Thr Leu Arg Tyr Arg Phe      73623  AGC TTC CCC CAT TCC AAG GTG GAG CTG TTG GCC TTG CTG GAT GCG GGC 67074  Ser Phe Pro His Ser Lys Val Glu Leu Leu Ala Leu Leu Asp Ala Gly      89671  ACT CTC TAC ACC TTC AGG TAC CAC GAG TGG CCC CGA GGC CAC GCA CCC     71890  Thr Leu Tyr Thr Phe Arg Tyr His Glu Trp Pro Arg Gly His Ala Pro     105719  ACA GAC TAT GCC CGC TGG CGG GAG GCT CAG GCC CCG TAC CGT GTG CAA     766106  Thr Asp Tyr Ala Arg Trp Arg Glu Ala Gln Ala Pro Tyr Arg Val Gln     121767  TGG GCG GCC AAC TAT GAA CCC TAC GTG GTG GTG CCA CGA GAC TGT CCC     814122  Trp Ala Ala Asn Tyr Glu Pro Tyr Val Val Val Pro Arg Asp Cys Pro     137 815  CGC TAT GAT CCT CGC TTT GTG GGC TTC GGC TGG AAC AAA GTG GCC CAC     862138  Arg Tyr Asp Pro Arg Phe Val Gly Phe Gly Trp Asn Lys Val Ala His     153863  ATT GTG GAG CTG GAT GCC CAG GAA TAT GAG CTC CTG GTG CTG CCC GAG     910154  Ile Val Glu Leu Asp Ala Gln Glu Tyr Glu Leu Leu Val Leu Pro Glu     169911  GCC TTC ACC ATC CAT CTG CCC CAC GCT CCA AGC CTG GAC ATC TCC GCT     958170  Ala Phe Thr Ile His Leu Pro His Ala Pro Ser Leu Asp Ile Ser Ala     185959  TCC GCT CCA GCC CCA CCT ATC GTG ACT GCC TCC AGG CCC TCA AGG ACG    1006186  Ser Ala Pro Ala Pro Pro Ile Val Thr Ala Ser Arg Pro Ser Arg Thr     2011007  AAT TCC ACC AGG ACT TGT CCC GCC ACC ATG GGG CTG CTG CCC TCA AAT    1054202  Asn Ser Thr Arg Thr Cys Pro Ala Thr Met Gly Leu Leu Pro Ser Asn     2171055  ACC TCC CAG CCC TGC AGC AGC CCC AGA GCC CTG CCC GAG GCT GAG GCT    1102218  Thr Ser Gln Pro Cys Ser Ser Pro Arg Ala Leu Pro Glu Ala Glu Ala     2331103  GGG CCG GCG CTG CCC CTC ATC TTA GCA TTG GGC AGA CAC CAG GGC AAC    1150234  Gly Pro Ala Leu Pro Leu Ile Leu Ala Leu Gly Arg His Gln Gly Asn     2491151  CTG CCC TCC GCC ATC CCT GCT ATT TAA ATT ATT TAA GGT CTC TGG GAA    1198250  Leu Pro Ser Ala Ile Pro Ala Ile ***                                 2581199  GGG CTG GGG CAG AGC ATC TGT GGG GTG GGG TCT TCC CCT TGC TGC TAT    12461247  TGT ATG GCT GGG GAC TGG TCT CTC TCT GCC CCA GCC AGT TTG GGG CTG    12941295  GTT CCC CCA TCT TGA ATT GTT TAT CCC TTT TTC ATA ATT AAA GTT TTA    13421343  AAA CAT AAA AAA AAA AAA AAA AAA AAA AAA A 1373D. Blastp result Query=PP5656 protein (257 amino acids)>SP_IN:Q21389 Q21389 caenorhabditis elegans.k09c8.4 protein.11/1999

Length = 622 amino acids Score = 104bits (256), predicted value = 8e-22 identity = 65/166 (39%), similarity = 93/166 (55%), gap = 17/166 (10%) Query:26 YVFLSDIDFLPAYSLYDYLRASIEQLGLGSRRKAALVVPAFET----LRYRFSFPHSKVE 81

      Y+ ++D+DF+    L DY    I+Q G   ++K  LV+PA E     LR   S   S+ +Sbjct:427 YILMTDVDFVV---LGDY-GTIIDQTG-NLKQKEVLVIPALEMTYPQLRLNLSNFLSRKD 481Query:82  LLA--LLDAGTLYTFRYHEWPRGHAPTDYARWREAQAPY----RVQWAANYEPYVVVPRD 135

      L+   LL+  T+ TFR   WP  H PT+ ++W ++   Y     V +  NYEPY V+ ++Sbjct:482 LVIEHLLNK-TIQTFRETIWPSSHVPTNISKWIKSNRTYMVAQNVNYEKNYEPYFVIKKE 540Query:136 -CPRYDPRFVGFGWNKVAHIVELDAQEYELLVLPEAFTIHLPHAPS 180

CP YD RF GFGWNKV H+++L Y+ LV P +F IH H SSbjct:541 ECPFYDQRFGGFGWNKVTHVMQLKMMNYKFLVSPTSFMIHQNHNAS 586

All documents mentioned in this application are incorporated by reference in this application as if each were individually incorporated by reference. In addition, it should be understood that after reading the above teaching content of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the appended claims of the present application.

Claims (11)

1. isolating people's albumen with cancer suppressing function, it is characterized in that it comprises the polypeptide of the aminoacid sequence with the group of being selected from down: SEQ ID NO:2, SEQ ID NO:5, SEQ ID NO:8, SEQ ID NO:11, SEQ ID NO:14, SEQID NO:17, SEQ ID NO:20, SEQ ID NO:23, SEQ ID NO:26;

Or its conservative property variation polypeptide or its active fragments or its reactive derivative.

2. polypeptide as claimed in claim 1, it is characterized in that this polypeptide is the polypeptide with aminoacid sequence of the group of being selected from down: SEQ ID NO:2, SEQ ID NO:5, SEQ ID NO:8, SEQ ID NO:11, SEQ ID NO:14, SEQ ID NO:17, SEQ ID NO:20, SEQ ID NO:23, SEQ ID NO:26.

3. isolating polynucleotide is characterized in that, it comprises a nucleotide sequence, and this nucleotide sequence is shown at least 85% homogeny with a kind of nucleotides sequence that is selected from down group:

(a) coding is as the polynucleotide of polypeptide as described in claim 1 and 2;

(b) with polynucleotide (a) complementary polynucleotide.

4. polynucleotide as claimed in claim 3, it is characterized in that the polypeptide of this polynucleotide encoding has the aminoacid sequence of the group of being selected from down: SEQ ID NO:2, SEQ ID NO:5, SEQ ID NO:8, SEQ ID NO:11, SEQ ID NO:14, SEQ ID NO:17, SEQ ID NO:20, SEQ ID NO:23, SEQ ID NO:26.

5. polynucleotide as claimed in claim 3 is characterized in that, the sequence of these polynucleotide is selected from down group:

Coding region sequence or the full length sequence of SEQ ID NO:3, SEQ ID NO:6, SEQ ID NO:9, SEQ ID NO:12, SEQ ID NO:15, SEQ ID NO:18, SEQ ID NO:21, SEQ ID NO:24, SEQ ID NO:27.

6. a carrier is characterized in that, it contains the described polynucleotide of claim 3.

7. a genetically engineered host cell is characterized in that, it is a kind of host cell that is selected from down group:

(a) host cell that transforms or transduce with the described carrier of claim 6;

(b) host cell that transforms or transduce with the described polynucleotide of claim 3.

8. the preparation method of the polypeptide of the people's protein-active with cancer suppressing function is characterized in that this method comprises:

(a) have under the proteic condition of people of cancer suppressing function suitable the expression, cultivate the described host cell of claim 7;

(b) from culture, isolate the polypeptide of people's protein-active with cancer suppressing function.

9. energy and the described people's protein-specific bonded antibody of claim 1 with cancer suppressing function.

10. nucleic acid molecule, it contains a successive 10-800 Nucleotide in the described polynucleotide of claim 3.

11. a pharmaceutical composition is characterized in that, it contains the described polypeptide of claim 1 and the pharmaceutically acceptable carrier of safe and effective amount.