CN1351079A - Human protein with cancer cell growth suppressing function and its coding sequence - Google Patents

Abstract

A novel human protein with cancer suppressing function, the polynucleotide for coding it, the process for preparing said polypeptide by recombination, the application of said polypeptide in treating diseases such as cancer, etc, the antagon of said polypeptide and its medical action, and the application of said polynucleotide are disclosed.

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; or its conservative variant polypeptide, or its active fragment, or its active derivative.

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.

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. 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 , the coding region sequence or full-length sequence of SEQ ID NO: 21.

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 that can be used for detection, which contains 10 consecutive nucleotides to the full-length nucleotides of the above-mentioned polynucleotide, preferably it contains continuous about 10-800 nucleotides.

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 PP8153 protein (in this application, the protein is named using its clone number) as an example, the coding region sequence encoding the mature polypeptide can be the same as the coding region sequence shown in SEQ ID NO: 3 or a degenerate variant. As used herein, "degenerate variant" in the present invention refers to a nucleic acid sequence that encodes a protein with SEQ ID NO: 2, but differs from the sequence of the coding region shown in SEQ ID NO: 3. Taking the PP8332 protein (in this application, the protein is named using its clone number) as an example, the coding region sequence encoding the mature polypeptide can be the same as the coding region sequence shown in SEQ ID NO: 6 or a degenerate variant. 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" refers 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, 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 (taking PP8153 protein as an example) 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). And constructing a cDNA library is also a common method (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 nucleotide sequence of the gene of the present invention obtained as described above, or various DNA fragments, etc., can be determined 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 (Science, 1984; 224: 1431), the polynucleotide sequence of the present invention can be used to express or produce recombinant protein polypeptides with tumor suppressor function. Generally speaking, there are the following steps:

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

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

(3). Isolate and purify protein 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. Vectors applicable in the present invention include, but are not limited to: T7-based expression vectors (Rosenberg, et al. Gene, 1987, 56: 125) expressed in bacteria; 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, LTRs of retroviruses and other promoters known to control gene expression 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 polypeptides of the invention can be used in combination with other therapeutic compounds.

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 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, for example, 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's instructions suggested conditions.

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

SP2114a was obtained from a liver cDNA library purchased from GIBCO BRL (catalogue number: 10422-012), and PP8153, PP8332, PP9177, PP9445, PP10199 and PP10226 were obtained by constructing a human placenta cDNA library by conventional methods. The placental tissue of 3, 6, and 10 months old was taken, and the total RNA was extracted with Trizol reagent (GIBCOBRL company) according to the manufacturer's instructions, and the mRNA was extracted with the 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. XL 10-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 using 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 incubate at 37°C for 24 hours. Change 100 μl whole culture medium for each well, 37°C for 24 hours, change 100 μl whole culture medium containing G418, 37°C for 24-48 hours, change the culture medium with different concentrations of G418 while observing. After about 2 to 3 times, until the microscopic examination of the cells shows 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) PP8153PP8332PP9177PP9445PP10199PP10226SP2114a 2 0 18 3 516 11 156 1 95 2 26 1 34 6 5 26 29 3012 13 1548 38 3548 38 3548 38 3548 38 3538 42 40

The dideoxy termination method was used for the above cDNA clones, 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, but the full-length cDNA sequence was still not obtained, primers were designed, and sequencing was performed again until the full-length sequence (SEQ ID NO: 1, 4, 7, 10, 13, 16 , 19).

Embodiment 2: PCR obtains full-length gene 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). Use MMLV-RT-Superscript II (GIBCO BRL) and reverse transcriptase to perform reverse transcription reaction at 42°C to obtain placental cDNA. Using the transmutation primers of each gene (as shown in the table below), follow 97°C for 3 minutes, 1 cycle; 94°C for 30 seconds → 60°C for 30 seconds → 72°C for 1 minute, a total of 35 cycles; 72°C for 10 minutes, 1 cycle. PCR amplification was carried out for one cycle to obtain the amplified products of each protein gene containing the complete open reading frame sequence. 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. (Note: For SP2114a, a liver cDNA library (catalogue number: 10422-012) purchased from GIBCO BRL company can be used as a template).

gene specific primers clone name Specific primer 1 (5'→3') Specific primer 2 (5'→3') PP8153PP8332PP9177PP9445PP10199PP10226SP2114a CGGAGGTTCTAGTGTCGGAGTTTCAGACCTGTTCCAAGGGCAGAGGAGCATCCCGTCTACACCAAATGAGGGACATGGAAGCAGTACTCCATGGTGCAGAAATGCACCCTGTTTTGAGAGACCACTTCCCACCAGAGACACA GGACTGCTACCCATCCTGAACTCTGCCTCCACTCACACTGGCTAGCCAGCTCTGTGGAGTCCAAGCCTGACTCTCTTTGCGTTTGCTCCCAGCTGTCTTCAGATCAACTTGAGGCCAGGAGCAAGGTTTCTTCAACTGGC Embodiment 3: cDNA clone sequence analysis

1. PP8153

A：核苷酸序列(SEQ ID NO：1)长度：2349bp1    GTGGAAGTAG  AAGGCGGTGG  CTGAGGCGGT  TCCGGAGGTT  CTAGTGTCGG  AGTTGGGTGC61    AGGCAGGTGC  CATGGGCCCG  CTTGAGGCAC  ACTGAGGGGA  CGCGGGGCTG  GGCCATGGCC121    GGCGCTCGGG  CCGCCGCCGC  CGCTGCCTCG  GCGGGGTCCT  CGGCCTCTTC  AGGCAACCAG181    CCGCCTCAGG  AGCTGGGGCT  TGGGGAGCTG  CTGGAGGAGT  TCTCCCGGAC  TCAGTACCGG241    GCCAAGGATG  GCAGCGGGAC  CGGCGGCTCT  AAGGTTGAGC  GCATTGAGAA  GAGATGTCTG301    GAGCTGTTTG  GCCGAGACTA  CTGTTTCAGC  GTGATTCCAA ACACGAATGG  GGATATCTGT361    GGCCACTATC  CCCGGCACAT  CGTGTTCCTG  GAGTATGAGA  GTTCTGAGAA  GGAGAAAGAC421    ACGTTTGAGA  GTACCGTACA  GGTGAGCAAG  TTGCAAGACC  TCATCCACCG  CAGCAAGATG481    GCCCGGTGCA  GAGGACGGTT  TGTCTGCCCA  GTAATCCTGT  TCAAGGGCAA  GCACATTTGC541    AGGTCGGCCA  CACTGGCTGG  ATGGGGAGAG  CTGTATGGAC  GCTCAGGCTA  CAACTATTTT601    TTCTCAGGGG  GTGCAGATGA  TGCCTGGGCA  GATGTGGAGG  ACGTCACGGA  GGAGGACTGT661    GCTCTTCGAA  GTGGTGACAC  GCATCTTTTT  GATAAGGTCA  GAGGCTATGA  CATCAAGCTG721    CTTCGATACC  TGTCAGTCAA  ATACATCTGT  GACCTGATGG  TGGAGAACAA  GAAGGTGAAG781    TTTGGCATGA  ATGTAACCTC  CTCTGAGAAG  GTGGACAAAG  CCCAGCGCTA  TGCCCACTTC841 ACTCTCCTCT  CCATCCCGTA  TCCAGGCTGT  GAATTTTTCA  AGGAATATAA  AGATCGGGAT901    TACATGGCAG  AAGGGCTCAT  ATTTAACTGG  AAGCAGGACT  ACGTTGATGC  CCCATTGAGC961    ATCCCCGACT  TCCTGACTCA  CTCTCTGAAC  ATTGACTGGA  GCCAGTATCA  GTGTTGGGAT1021    CTGGTGCAAC  AAACACAAAA  CTACCTGAAG  CTGCTGCTTT  CCTTAGTTAA  CAGTGATGAT1081    GACAGCGGGC  TGCTGGTACA  CTGTATCTCA  GGCTGGGATC  GGACCCCCCT  CTTCATCTCC1141    CTCCTGCGCC  TTTCCTTGTG  GGCTGATGGG  CTCATCCACA  CGTCCCTGAA  GCCCACTGAG1201    ATCCTCTACC  TCACTGTGGC  CTATGACTGG  TTCCTCTTCG  GGCACATGTT  GGTAGATCGG1261    CTCAGCAAAG  GGGAGGAGAT  TTTCTTCTTC  TGCTTCAATT  TTTTGAAGCA  TATTACCTCC1321    GAGGAGTTCT  CTGCTCTGAA GACCCAGAGG  AGGAAGAGTT  TGCCAGCCCG  GGATGGAGGC1381    TTCACCCTGG  AAGACATCTG  CATGCTGAGA  CGAAAGGACC  GTGGCAGCAC  CACCAGCCTT1441    GGCAGCGACT  TCTCCCTGGT  CATGGAGAGT  TCCCCAGGAG  CCACTGGGAG  CTTCACCTAT1501    GAAGGCCGTG  GAGCTGGTCC  CAGCAGGAGC  GCCAACTCAG  GCAGCTTGAA  GGAAGAGCCA1561    CTCATCCTCT  CCACAGAGTG  TCCTCTGGAA  CCGGCCACAA  CCCTCAGAGG  ACCGCTTGCC1621    TTCCCAGCAG  GGGCTGGCGG  AAGCCAGGTC  TTCCAGCTCC  TCTTCCTCAA  ACCATTCTGA1681    TAACTTTTTC  AGGATGGGTA  GCAGTCCCCT  GGAGGTCCCC  AAACCCAGGC  TTGCAGCCCT1741    GAGTGATCGA  GAGACTCGGC  TGCAGGAGGT  GCGCTCAGCC  TTCTTGGCTG  CGTACAGCAG1801    CACAGTGGGG  CTTCGGGCAG  TAGCCCCCAG  TCCTTCCGGT GCCATCGGGG  GCCTGCTGGA1861    GCAATTTGCC  CGTGGTGTTG  GACTCCGGAG  CATCAGCAGC  AATGCCTTGT  GAAGAAGCCA1921    GCCCATGACA  TTTTCCTGCT  CCTCTCTCAG  CTGAGCCCTT  AGCAGAGAAT  CAAAGCCATG1981    CCTGGCCGAA  GGGGTACTTC  CAGGTCAGGG  GAAATTTCAG  TCCCCCATCT  CCATCATGAA2041    CATGGCAGCC  CCAAAGCTGA  GCAAGGCCAA  AGACAGGGTT  TTCCAACCCC  CAGCCTCTTG2101    ACTGGTGACC  ACCACCCCTT  CTTGTCACTG  TCTCCCACCC  ACCCCATCTT  TGCTGGGATT2161    CCCATCAACT  CTCAGAACTG  TGTGGGGTTT  CCCTGGGGCC  TTGTGGAAGC  CATGACTTCA2221    CAAAGACCCT  ACCTGTCAGT  TCTTGTTTCT  GGGGAGGAGG  GATCACCTGC  ACTGAGAATG2281    AGGCAGTTTG  ACACAGATCA  CAAAATAAAA  TCAAAGTCTT  TTTGAATAGC  CAAAAAAAAA2341 AAAAAAAAAAB：氨基酸序列(SEQ ID NO：2)长度：521个氨基酸1    MAGARAAAAA  ASAGSSASSG  NQPPQELGLG  ELLEEFSRTQ  YRAKDGSGTG  GSKVERIEKR61    CLELFGRDYC  FSVIPNTNGD  ICGHYPRHIV  FLEYESSEKE  KDTFESTVQV  SKLQDLIHRS121    KMARCRGRFV  CPVILFKGKH  ICRSATLAGW  GELYGRSGYN  YFFSGGADDA  WADVEDVTEE181    DCALRSGDTH  LFDKVRGYDI  KLLRYLSVKY  ICDLMVENKK  VKFGMNVTSS  EKVDKAQRYA241    DFTLLSIPYP  GCEFFKEYKD  RDYMAEGLIF  NWKQDYVDAP  LSIPDFLTHS  LNIDWSQYQC301    WDLVQQTQNY  LKLLLSLVNS  DDDSGLLVHC ISGWDRTPLF  ISLLRLSLWA  DGLIHTSLKP361    TEILYLTVAY  DWFLFGHMLV  DRLSKGEEIF  FFCFNFLKHI  TSEEFSALKT  QRRKSLPARD421    GGFTLEDICM  LRRKDRGSTT  SLGSDFSLVM  ESSPGATGSF  TYEGRGAGPS  RSANSGSLKE481    EPLILSTECP  LEPATTLRGP  LAFPAGAGGS  QVFQLLFLKP  FC.核苷酸及氨基酸组合序列(SEQ ID NO：3)克隆号：PP8153起始编码子：115 ATG    终止编码子： 1678 TGA蛋白质分子量：58350.051  GTG GAA GTA GAA GGC GGT GGC TGA GGC GGT TCC GGA GGT TCT AGT GTC      4849  GGA GTT GGG TGC AGG CAG GTG CCA TGG GCC CGC TTG AGG CAC ACT GAG      9697  GGG ACG CGG GGC TGG GCC ATG GCC GGC GCT CGG GCC GCC GCC GCC GCT 1441 MET Ala Gly Ala Ala Ala Ala Ala Ala Ala 10145 GCC TCG GGG GGG TCC TCC TCA GGC AAC CCG CCG CCG CCG CCG CCG CCG CCG CCG CCG CCG CCG CCG CCG CCG CCG CCG CCG CCG CCG CCG CCG CCG CCG CCG CCG CCG CCG CCG CCG CCG CCG CCG CCG CCG CCG CCG CCG CCG CAG Ala GLY SELA SELA Ser Ser Ser Ser Ser Ser Sera -G's G Gl that Fly PLT that PLC that PLC that Glu      26193  CTG GGG CTT GGG GAG CTG CTG GAG GAG TTC TCC CGG ACT CAG TAC CGG     24027  Leu Gly Leu Gly Glu Leu Leu Glu Glu Phe Ser Arg Thr Gln Tyr Arg      42241  GCC AAG GAT GGC AGC GGG ACC GGC GGC TCT AAG GTT GAG CGC ATT GAG     288  43  Ala Lys Asp Gly Ser Gly Thr Gly Gly Ser Lys Val Glu Arg Ile Glu       58289  AAG AGA TGT CTG GAG CTG TTT GGC CGA GAC TAC TGT TTC AGC GTG ATT      33659  Lys Arg Cys Leu Glu Leu Phe Gly Arg Asp Tyr Cys PHE Ser Val Ile 7437 CCA ACG AAT GGG GGG GAT ATC TGT GGC CAC CCC CCC CAC GTG 38475 Pro Asn THR Asp Ile Cyr Pro ARG HIS Ile Vag Gag Gag Gag Tat Gag Tat Gag Tat Gag Tat Gag Tat Gag Tat Gag Tat Gag Ag Tat Gag Tat Gag Tat Gag Tat Gag Tat Gag Tat Gag Tat Gag Tat Gag Tat Gag Tat Gag Tat Gag Tat Gag Tat Gag Tat Gag Tat Gag Tat Gag Tat Gag Tat Gag Tat Gag Tat Gag Tat Gag Tat Gag Tat Gag Tat Gag Tat Gag Tat Gag Tat Gag Tat Gag Tat Gag Tat Gag Agg GAC ACG TTT GAG AGT      43291  Phe Leu Glu Tyr Glu Ser Ser Glu Lys Glu Lys Asp Thr Phe Glu Ser      106433  ACC GTA CAG GTG AGC AAG TTG CAA GAC CTC ATC CAC CGC AGC AAG ATG      480107  Thr Val Gln Val Ser Lys Leu Gln Asp Leu Ile His Arg Ser Lys Met      122481  GCC CGG TGC AGA GGA CGG TTT GTC TGC CCA GTA ATC CTG TTC AAG GGC      528123  Ala Arg Cys Arg Gly Arg Phe Val Cys Pro Val Ile Leu Phe Lys Gly      138529  AAG CAC ATT TGC AGG TCG GCC ACA CTG GCT GGA TGG GGA GAG CTG TAT      576139  Lys His Ile Cys Arg Ser Ala Thr Leu Ala Gly Trp Gly Glu Leu Tyr      154577  GGA CGC TCA GGC TAC AAC TAT TTT TTC TCA GGG GGT GCA GAT GAT GCC      624155  Gly Arg Ser Gly Tyr Asn Tyr Phe Phe Ser Gly Gly Ala Asp Asp Ala      170625  TGG GCA GAT GTG GAG GAC GTC ACG GAG GAG GAC TGT GCT CTT CGA AGT      672171  Trp Ala Asp Val Glu Asp Val Thr Glu Glu Asp Cys Ala Leu Arg Ser      186673  GGT GAC ACG CAT CTT TTT GAT AAG GTC AGA GGC TAT GAC ATC AAG CTG      720187  Gly Asp Thr His Leu Phe Asp Lys Val Arg Gly Tyr Asp Ile Lys Leu      202721  CTT CGA TAC CTG TCA GTC AAA TAC ATC TGT GAC CTG ATG GTG GAG AAC      768203  Leu Arg Tyr Leu Ser Val Lys Tyr Ile Cys Asp Leu Met Val Glu Asn      218769  AAG AAG GTG AAG TTT GGC ATG AAT GTA ACC TCC TCT GAG AAG GTG GAC      816219  Lys Lys Val Lys Phe Gly Met Asn Val Thr Ser Ser Glu Lys Val Asp      234817  AAA GCC CAG CGC TAT GCC GAC TTC ACT CTC CTC TCC ATC CCG TAT CCA      864235  Lys Ala Gln Arg Tyr Ala Asp Phe Thr Leu Leu Ser Ile Pro Tyr Pro      250865  GGC TGT GAA TTT TTC AAG GAA TAT AAA GAT CGG GAT TAC ATG GCA GAA      912251  Gly Cys Glu Phe Phe Lys Glu Tyr Lys Asp Arg Asp Tyr Met Ala Glu      266913  GGG CTC ATA TTT AAC TGG AAG CAG GAC TAC GTT GAT GCC CCA TTG AGC      960267  Gly Leu Ile Phe Asn Trp Lys Gln Asp Tyr Val Asp Ala Pro Leu Ser      282961  ATC CCC GAC TTC CTG ACT CAC TCT CTG AAC ATT GAC TGG AGC CAG TAT     1008283  Ile Pro Asp Phe Leu Thr His Ser Leu Asn Ile Asp Trp Ser Gln Tyr      2981009  CAG TGT TGG GAT CTG GTG CAA CAA ACA CAA AAC TAC CTG AAG CTG CTG     1056299  Gln Cys Trp Asp Leu Val Gln Gln Thr Gln Asn Tyr Leu Lys Leu Leu      3141057  CTT TCC TTA GTT AAC AGT GAT GAT GAC AGC GGG CTG CTG GTA CAC TGT     1104315  Leu Ser Leu Val Asn Ser Asp Asp Asp Ser Gly Leu Leu Val His Cys      3301105  ATC TCA GGC TGG GAT CGG ACC CCC CTC TTC ATC TCC CTC CTG CGC CTT    1152331  Ile Ser Gly Trp Asp Arg Thr Pro Leu Phe Ile Ser Leu Leu Arg Leu     3461153  TCC TTG TGG GCT GAT GGG CTC ATC CAC ACG TCC CTG AAG CCC ACT GAG    1200347 Ser Leu Trp Ala Asp Gly Leu Ile His Thr Ser Leu Lys Pro Thr Glu     3621201  ATC CTC TAC CTC ACT GTG GCC TAT GAC TGG TTC CTC TTC GGG CAC ATG    1248363  Ile Leu Tyr Leu Thr Val Ala Tyr Asp Trp Phe Leu Phe Gly His Met 3781249  TTG GTA GAT CGG CTC AGC AAA GGG GAG GAG ATT TTC TTC TTC TGC TTC    1296379  Leu Val Asp Arg Leu Ser Lys Gly Glu Glu Ile Phe Phe Phe Cys Phe     3941297  AAT TTT TTG AAG CAT ATT ACC TCC GAG GAG TTC TCT GCT CTG AAG ACC    1344395  Asn Phe Leu Lys His Ile Thr Ser Glu Glu Phe Ser Ala Leu Lys Thr     4101345  CAG AGG AGG AAG AGT TTG CCA GCC CGG GAT GGA GGC TTC ACC CTG GAA    1392411  Gln Arg Arg Lys Ser Leu Pro Ala Arg Asp Gly Gly Phe Thr Leu Glu     4261393  GAC ATC TGC ATG CTG AGA CGA AAG GAC CGT GGC AGC ACC ACC AGC CTT    1440427  Asp Ile Cys Met Leu Arg Arg Lys Asp Arg Gly Ser Thr Thr Ser Leu     4421441  GGC AGC GAC TTC TCC CTG GTC ATG GAG AGT TCC CCA GGA GCC ACT GGG    1488443  Gly Ser Asp Phe Ser Leu Val Met Glu Ser Ser Pro Gly Ala Thr Gly     4581489  AGC TTC ACC TAT GAA GGC CGT GGA GCT GGT CCC AGC AGG AGC GCC AAC    1536459  Ser Phe Thr Tyr Glu Gly Arg Gly Ala Gly Pro Ser Arg Ser Ala Asn     4741537  TCA GGC AGC TTG AAG GAA GAG CCA CTC ATC CTC TCC ACA GAG TGT CCT    1584475  Ser Gly Ser Leu Lys Glu Glu Pro Leu Ile Leu Ser Thr Glu Cys Pro     4901585  CTG GAA CCG GCC ACA ACC CTC AGA GGA CCG CTT GCC TTC CCA GCA GGG    1632491  Leu Glu Pro Ala Thr Thr Leu Arg Gly Pro Leu Ala Phe Pro Ala Gly     5061633  GCT GGC GGA AGC CAG GTC TTC CAG CTC CTC TTC CTC AAA CCA TTC TGA    1680507  Ala Gly Gly Ser Gln Val Phe Gln Leu Leu Phe Leu Lys Pro Phe ***     5221681  TAA CTT TTT CAG GAT GGG TAG CAG TCC CCT GGA GGT CCC CAA ACC CAG    17281729  GCT TGC AGC CCT GAG TGA TCG AGA GAC TCG GCT GCA GGA GGT GCG CTC    17761777  AGC CTT CTT GGC TGC GTA CAG CAG CAC AGT GGG GCT TCG GGC AGT AGC    18241825  CCC CAG TCC TTC CGG TGC CAT CGG GGG CCT GCT GGA GCA ATT TGC CCG    18721873  TGG TGT TGG ACT CCG GAG CAT CAG CAG CAA TGC CTT GTG AAG AAG CCA    19201921  GCC CAT GAC ATT TTC CTG CTC CTC TCT CAG CTG AGC CCT TAG CAG AGA    19681969  ATC AAA GCC ATG CCT GGC CGA AGG GGT ACT TCC AGG TCA GGG GAA ATT    20162017  TCA GTC CCC CAT CTC CAT CAT GAA CAT GGC AGC CCC AAA GCT GAG CAA    20642065  GGC CAA AGA CAG GGT TTT CCA ACC CCC AGC CTC TTG ACT GGT GAC CAC    21122113  CAC CCC TTC TTG TCA CTG TCT CCC ACC CAC CCC ATC TTT GCT GGG ATT    21602161  CCC ATC AAC TCT CAG AAC TGT GTG GGG TTT CCC TGG GGC CTT GTG GAA    22082209  GCC ATG ACT TCA CAA AGA CCC TAC CTG TCA GTT CTT GTT TCT GGG GAG    22562257  GAG GGA TCA CCT GCA CTG AGA ATG AGG CAG TTT GAC ACA GAT CAC AAA    23042305  ATA AAA TCA AAG TCT TTT TGA ATA GCC AAA AAA AAA AAA AAA AAA        23492.PP8332A：核苷酸序列(SEQ ID NO：4)长度：1771bp1    GCCTGGGGCG  TCCCCGCGAA  GCCTGGGCCT  GTCAGGCGGT  TCCGTCCGGG  TCTCGGCCAC61    CGTCGAGTTC  CGTCGAGTTC  CGTCCCGGCC  CTGCTCACAG  CAGCGCCCTC  GGAGCGCCCA121    GCACCTGCGG  CCGGCCAGGC  AGCGCGATCC  TGCGGCGTCT  GGCCATCCCG  AATGCTATGG181    CCGCCGTCGC  CGTCTTGCGG  GCCTTCGGGG  CAAGTGGGCC  CATGTGTCTC  CGGCGCGGCC241    CCTGGGCCCA  GCTCCCCGCC  CGCTTCTGCA  GCCGGGACCC  GGCCGGGGCG  GGGCGGCGGG301    AGTCGGAGCC  GCGGCCCACC  AGCGCGCGGC  AGCTGGACGG  CATAAGGAAC  ATCGTCTTGA361    GCAATCCCAA GAAGAGGAAC  ACGTTGTCAC  TTGCAATGCT  GAAATCTCTC  CAAAGTGACA421    TTCTTCATGA  CGCTGACAGC  AACGATCTGA  AAGTCATTAT  CATCTCGGCT  GAGGGGCCTG481    TGTTTTCTTC  TGGGCATGAC  TTAAAGGAGC  TGACAGAGGA  GCAAGGCCGT  GATTACCATG541    CCGAAGTATT  TCAGACCTGT  TCCAAGGGTC  TCGCTCTGTC  GCCCAGGCTG  GATTACAGTG601    GCATGATCTC  GGCTCACTGC  AACCTCTGCC  TCCCGGGTTC  AAGCAATTCT  CCTGCCTCAG661    CCTCCTGAGT  AGCTGGGACT  ACAGGTCATG  ATGCACATCC  GGAACCACCC  CGTCCCCGTC721    ATTGCCATGG  TCAATGGCCT  GGCCACGGCT  GCCGGCTGTC  AACTGGTTGC  CAGCTGCGAC781    ATTGCCGTGG  CGAGCGACAA  GTCCTCTTTT  GCCACTCCTG  GGGTGAACGT  CGGGCTCTTC841    TGTTCTACCC  CTGGGGTTGC  CTTGGCAAGA GCAGTGCCTA  GAAAGGTGGC  CTTGGAGATG901    CTCTTTACTG  GTGAGCCCAT  TTCTGCCCAG  GAGGCCCTGC  TCCACGGGCT  GCTTAGCAAG961    GTGGTGCCAG  AGGCGGAGCT  GCAGGAGGAG  ACCATGCGGA  TCGCTAGGAA  GATCGCATCG1021    CTGAGCCGTC  CGGTGGTGTC  CCTGGGCAAA  GCCACCTTCT  ACAAGCAGCT  GCCCCAGGAC1081    CTGGGGACGG  CTTACTACCT  CACCTCCCAG  GCCATGGTGG  ACAACCTGGC  CCTGCGGGAC1141    GGGCAGGAGG  GCATCACGGC  CTTCCTCCAG  AAGAGAAAAC  CTGTCTGGTC  ACACGAGCCA1201    GTGTGAGTGG  AGGCAGAGGA  GTGAGGCCCA  CGGGCAGCGC  CCAGGAGCCC  ACCTTCCCCT1261    CTGGCCCAGC  CACCACTGCC  TCTCAGCTTC  AACAGGTGAC  AGGCTGCTTT  CGTGACTTGA1321    TATTGGTGTC  ATAGCATTTG  GCCTACATTA  AAAGCCACAA  TTTCATGGGG AAAGGACAAA1381    ATGGAGAGTG  ACTGAGGTGC  TGACCTCAGT  GCAAGGCTGG  TGAACCCTGC  AGCGGGCCAG1441    CTATGGTGGG  AAGCCTGGCA  TTTGGGGTGC  TCCTTGCAAC  GTCTTAAGCA  AGCGACCCCC1501    CTGACATAGC  AAAAGGTGGC  AACCCATGGA  GGCAGAAAGA  AGGACGCCAG  CCTGACCCTT1561    ATCTTGAAAC  GTCCTAAGCA  GAGTTAATCC  TGGCTGCTCA  GGAGAGGCGA  CACATTTCAA1621    ATCTCCACGA  GATATTCTCC  ACACAGAAAA  TCTTCTTGAT  TCTATAGAGA  CTTAATCATG1681    CCTATGGCTT  TGAATAATCT  TATGTGATTT  AAATAAATTA  AATCTTTATA  GAGACTGGAA1741    AAAAAAAAAA  AAAAAAAAAA  AAAAAAAAAA  A

B：氨基酸序列(SEQ ID NO：5)长度：172个氨基酸1    MMHIRNHPVP  VIAMVNGLAT  AAGCQLVASC  DIAVASDKSS  FATPGVNVGL  FCSTPGVALA61    RAVPRKVALE  MLFTGEPISA  QEALLHGLLS  KVVPEAELQE  ETMRIARKIA  SLSRPVVSLG121    KATFYKQLPQ  DLGTAYYLTS  QAMVDNLALR  DGQEGITAFL  QKRKPVWSHE  PV

C. Nucleotide and amino acid combination sequence (SEQ ID NO: 6)

Clone number: PP8332

Start code: 688 ATG End code: 1204 TGA

蛋白质分子量：18409.481  GCC TGG GGC GTC CCC GCG AAG CCT GGG CCT GTC AGG CGG TTC CGT CCG     4849  GGT CTC GGC CAC CGT CGA GTT CCG TCG AGT TCC GTC CCG GCC CTG CTC     9697  ACA GCA GCG CCC TCG GAG CGC CCA GCA CCT GCG GCC GGC CAG GCA GCG    144145  CGA TCC TGC GGC GTC TGG CCA TCC CGA ATG CTA TGG CCG CCG TCG CCG    192193  TCT TGC GGG CCT TCG GGG CAA GTG GGC CCA TGT GTC TCC GGC GCG GCC    240241  CCT GGG CCC AGC TCC CCG CCC GCT TCT GCA GCC GGG ACC CGG CCG GGG    288289  CGG GGC GGC GGG AGT CGG AGC CGC GGC CCA CCA GCG CGC GGC AGC TGG    336 337  ACG GCA TAA GGA ACA TCG TCT TGA GCA ATC CCA AGA AGA GGA ACA CGT     384385  TGT CAC TTG CAA TGC TGA AAT CTC TCC AAA GTG ACA TTC TTC ATG ACG     432433  CTG ACA GCA ACG ATC TGA AAG TCA TTA TCA TCT CGG CTG AGG GGC CTG     480481  TGT TTT CTT CTG GGC ATG ACT TAA AGG AGC TGA CAG AGG AGC AAG GCC     528529  GTG ATT ACC ATG CCG AAG TAT TTC AGA CCT GTT CCA AGG GTC TCG CTC     576577  TGT CGC CCA GGC TGG ATT ACA GTG GCA TGA TCT CGG CTC ACT GCA ACC     624625  TCT GCC TCC CGG GTT CAA GCA ATT CTC CTG CCT CAG CCT CCT GAG TAG     672673  CTG GGA CTA CAG GTC ATG ATG CAC ATC CGG AAC CAC CCC GTC CCC GTC     7201                      Met Met His Ile Arg Asn His Pro Val Pro Val      11721  ATT GCC ATG GTC AAT GGC CTG GCC ACG GCT GCC GGC TGT CAA CTG GTT     76812  Ile Ala Met Val Asn Gly Leu Ala Thr Ala Ala Gly Cys Gln Leu Val      27769  GCC AGC TGC GAC ATT GCC GTG GCG AGC GAC AAG TCC TCT TTT GCC ACT     81628  Ala Ser Cys Asp Ile Ala Val Ala Ser Asp Lys Ser Ser Phe Ala Thr      43817  CCT GGG GTG AAC GTC GGG CTC TTC TGT TCT ACC CCT GGG GTT GCC TTG     86444  Pro Gly Val Asn Val Gly Leu Phe Cys Ser Thr Pro Gly Val Ala Leu      59865  GCA AGA GCA GTG CCT AGA AAG GTG GCC TTG GAG ATG CTC TTT ACT GGT     91260  Ala Arg Ala Val Pro Arg Lys Val Ala Leu Glu Met Leu Phe Thr Gly      75913  GAG CCC ATT TCT GCC CAG GAG GCC CTG CTC CAC GGG CTG CTT AGC AAG     96076  Glu Pro Ile Ser Ala Gln Glu Ala Leu Leu His Gly Leu Leu Ser Lys      91961  GTG GTG CCA GAG GCG GAG CTG CAG GAG GAG ACC ATG CGG ATC GCT AGG    100892  Val Val Pro Glu Ala Glu Leu Gln Glu Glu Thr Met Arg Ile Ala Arg     1071009  AAG ATC GCA TCG CTG AGC CGT CCG GTG GTG TCC CTG GGC AAA GCC ACC    1056108  Lys Ile Ala Ser Leu Ser Arg Pro Val Val Ser Leu Gly Lys Ala Thr     1231057  TTC TAC AAG CAG CTG CCC CAG GAC CTG GGG ACG GCT TAC TAC CTC ACC    1104124  Phe Tyr Lys Gln Leu Pro Gln Asp Leu Gly Thr Ala Tyr Tyr Leu Thr     1391105  TCC CAG GCC ATG GTG GAC AAC CTG GCC CTG CGG GAC GGG CAG GAG GGC    1152140  Ser Gln Ala Met Val Asp Asn Leu Ala Leu Arg Asp Gly Gln Glu Gly     1551153  ATC ACG GCC TTC CTC CAG AAG AGA AAA CCT GTC TGG TCA CAC GAG CCA    1200156  Ile Thr Ala Phe Leu Gln Lys Arg Lys Pro Val Trp Ser His Glu Pro     1711201  GTG TGA GTG GAG GCA GAG GAG TGA GGC CCA CGG GCA GCG CCC AGG AGC    1248172  Val ***                                                             1731249  CCA CCT TCC CCT CTG GCC CAG CCA CCA CTG CCT CTC AGC TTC AAC AGG 12961297  TGA CAG GCT GCT TTC GTG ACT TGA TAT TGG TGT CAT AGC ATT TGG CCT    13441345  ACA TTA AAA GCC ACA ATT TCA TGG GGA AAG GAC AAA ATG GAG AGT GAC    13921393  TGA GGT GCT GAC CTC AGT GCA AGG CTG GTG AAC CCT GCA GCG GGC CAG    14401441  CTA TGG TGG GAA GCC TGG CAT TTG GGG TGC TCC TTG CAA CGT CTT AAG    14881489  CAA GCG ACC CCC CTG ACA TAG CAA AAG GTG GCA ACC CAT GGA GGC AGA    15361537  AAG AAG GAC GCC AGC CTG ACC CTT ATC TTG AAA CGT CCT AAG CAG AGT    15841585  TAA TCC TGG CTG CTC AGG AGA GGC GAC ACA TTT CAA ATC TCC ACG AGA    16321633  TAT TCT CCA CAC AGA AAA TCT TCT TGA TTC TAT AGA GAC TTA ATC ATG    16801681 CCT ATG GCT TTG AAT AAT CTT ATG TGA TTT AAA TAA ATT AAA TCT TTA 17281729 TAG AGA CTG GAA AAA AAA AAA AAA AAA AAA AAA AAA AAA AAA A 1771

3. PP9177

A：核苷酸序列(SEQ ID NO：7)长度：2160bp1  GCGTCTGCCA  GCCGGCTTGG  CTAGCGCGCG  GCGGCCGTGG  CTAAGGCTGC  TACGAAGCGA61  GCTTGGGAGG  AGCAGCGGCC  TGCGGGGCAG  AGGAGCATCC  CGTCTACCAG  GTCCCAAGCG121  GCCGTGGCCC  GCGGGTCATG  GCCAAAGGAG  AAGGCGCCGA  GAGCGGCTCC  GCGGCGGGGC181  TGCTACCCAC  CAGCATCCTC  CAAAGCACTG  AACGCCCGGC  CCAGGTGAAG  AAAGAACCGA241  AAAAGAAGAA  ACAACAGTTG  TCTGTTTGCA  ACAAGCTTTG  CTATGCACTT  GGGGGAGCCC301  CCTACCAGGT  GACGGGCTGT  GCCCTGGGTT  TCTTCCTTCA GATCTACCTA  TTGGATGTGG361  CTCAGGTGGG  CCCTTTCTCT  GCCTCCATCA  TCCTGTTTGT  GGGCCGAGCC  TGGGATGCCA421  TCACAGACCC  CCTGGTGGGC  CTCTGCATCA  GCAAATCCCC  CTGGACCTGC  CTGGGTCGCC481  TTATGCCCTG  GATCATCTTC  TCCACGCCCC  TGGCCGTCAT  TGCCTACTTC  CTCATCTGGT541  TCGTGCCCGA  CTTCCCACAC  GGCCAGACCT  ATTGGTACCT  GCTTTTCTAT  TGCCTCTTTG601  AAACAATGGT  CACGTGTTTC  CATGTTCCCT  ACTCGGCTCT  CACCATGTTC  ATCAGCACCG661  AGCAGACTGA  GCGGGATTCT  GCCACCGCCT  ATCGGATGAC  TGTGGAAGTG  CTGGGCACAG721  TGCTGGGCAC  GGCGATCCAG  GGACAAATCG  TGGGCCAAGC  AGACACGCCT  TGTTTCCAGG781  ACCTCAATAG  CTCTACAGTA  GCTTCACAAA  GTGCCAACCA  TACACATGGC  ACCACCTCAC841 ACAGGGAAAC  GCAAAAGGCA  TACCTGCTGG  CAGCGGGGGT  CATTGTCTGT  ATCTATATAA901  TCTGTGCTGT  CATCCTGATC  CTGGGCGTGC  GGGAGCAGAG  AGAACCCTAT  GAAGCCCAGC961  AGTCTGAGCC  AATCGCCTAC  TTCCGGGGCC  TACGGCTGGT  CATGAGCCAC  GGCCCATACA1021  TCAAACTTAT  TACTGGCTTC  CTCTTCACCT  CCTTGGCTTT  CATGCTGGTG  GAGGGGAACT1081  TTGTCTTGTT  TTGCACCTAC  ACCTTGGGCT  TCCGCAATGA  ATTCCAGAAT  CTACTCCTGG1141  CCATCATGCT  CTCGGCCACT  TTAACCATTC  CCATCTGGCA  GTGGTTCTTG  ACCCGGTTTG1201  GCAAGAAGAC  AGCTGTATAT  GTTGGGATCT  CATCAGCAGT  GCCATTTCTC  ATCTTGGTGG1261  CCCTCATGGA  GAGTAACCTC  ATCATTACAT  ATGCGGTAGC  TGTGGCAGCT  GGCATCAGTG1321  TGGCAGCTGC  CTTCTTACTA CCCTGGTCCA  TGCTGCCTGA  TGTCATTGAC  GACTTCCATC1381  TGAAGCAGCC  CCACTTCCAT  GGAACCGAGC  CCATCTTCTT  CTCCTTCTAT  GTCTTCTTCA1441  CCAAGTTTGC  CTCTGGAGTG  TCACTGGGCA  TTTCTACCCT  CAGTCTGGAC  TTTGCAGGGT1501  ACCAGACCCG  TGGCTGCTCG  CAGCCGGAAC  GTGTCAAGTT  TACACTGAAC  ATGCTCGTGA1561  CCATGGCTCC  CATAGTTCTC  ATCCTGCTGG  GCCTGCTGCT  CTTCAAAATG  TACCCCATTG1621  ATGAGGAGAG  GCGGCGGCAG  AATAAGAAGG  CCCTGCAGGC  ACTGAGGGAC  GAGGCCAGCA1681  GCTCTGGCTG  CTCAGAAACA  GACTCCACAG  AGCTGGCTAG  CATCCTCTAG  GGCCCGCCAC1741  GTTGCCCGAA  GCCACCATGC  AGAAGGCCAC  AGAAGGGATC  AGGACCTGTC  TGCCGGCTTG1801  CTGAGCAGCT  GGACTGCAGG  TGCTAGGAAG  GGAACTGAAG ACTCAAGGAG  GTGGCCCAGG1861  ACACTTGCTG  TGCTCACTGT  GGGGCCGGCT  GCTCTGTGGC  CTCCTGCCTC  CCCTCTGCCT1921  GCCTGTGGGG  CCAAGCCCTG  GGGCTGCCAC  TGTGAATATG  CCAAGGACTG  ATCGGGCCTA1981  GCCCGGAACA  CTAATGTAGA  AACCTTTTTT  TTACAGAGCC  TAATTAATAA  CTTAATGACT2041  GTGTACATAG  CAATGTGTGT  GTATGTATAT  GTCTGTGAGC  TATTAATGTT  ATTAATTTTC2101  ATAAAAGCTG  GAAAGCAAAA  AAAAAAAAAA  AAAAAAAAAA  AAAAAAAAAA  AAAAAAAAAA

B：氨基酸序列(SEQ ID NO：8)  长度：530个氨基酸1   MAKGEGAESG  SAAGLLPTSI  LQSTERPAQV  KKEPKKKKQQ  LSVCNKLCYA  LGGAPYQVTG61   CALGFFLQIY  LLDVAQVGPF  SASIILFVGR  AWDAITDPLV  GLCISKSPWT  CLGRLMPWII121   FSTPLAVIAY  FLIWFVPDFP  HGQTYWYLLF  YCLFETMVTC  FHVPYSALTM  FISTEQTERD181   SATAYRMTVE  VLGTVLGTAI  QGQIVGQADT  PCFQDLNSST  VASQSANHTH  GTTSHRETQK241   AYLLAAGVIV  CIYIICAVIL  ILGVREQREP  YEAQQSEPIA  YFRGLRLVMS  HGPYIKLITG301   FLFTSLAFML  VEGNFVLFCT  YTLGFRNEFQ NLLLAIMLSA  TLTIPIWQWF  LTRFGKKTAV361   YVGISSAVPF  LILVALMESN  LIITYAVAVA  AGISVAAAFL  LPWSMLPDVI  DDFHLKQPHF421   HGTEPIFFSF  YVFFTKFASG  VSLGISTLSL  DFAGYQTRGC  SQPERVKFTL  NMLVTMAPIV481 LILLGLLLFK MYPIDEERRR QNKKALQALR DEASSSGCSE TDSTELASIL

C. Nucleotide and amino acid combination sequence (SEQ ID NO: 9)

Clone number: PP9177

Start code: 138 ATG End code: 1728 TAG

蛋白质分子量：58620.361   GC GTC TGC CAG CCG GCT TGG CTA GCG CGC GGC GGC CGT GGC TAA GGC      4748  TGC TAC GAA GCG AGC TTG GGA GGA GCA GCG GCC TGC GGG GCA GAG GAG      9596  CAT CCC GTC TAC CAG GTC CCA AGC GGC CGT GGC CCG CGG GTC ATG GCC     1431                                                          Met Ala       2144  AAA GGA GAA GGC GCC GAG AGC GGC TCC GCG GCG GGG CTG CTA CCC ACC     1913  Lys Gly Glu Gly Ala Glu Ser Gly Ser Ala Ala Gly Leu Leu Pro Thr      18192  AGC ATC CTC CAA AGC ACT GAA CGC CCG GCC CAG GTG AAG AAA GAA CCG     23919  Ser Ile Leu Gln Ser Thr Glu Arg Pro Ala Gln Val Lys Lys Glu Pro      34240  AAA AAG AAG AAA CAA CAG TTG TCT GTT TGC AAC AAG CTT TGC TAT GCA     28735  Lys Lys Lys Lys Gln Gln Leu Ser Val Cys Asn Lys Leu Cys Tyr Ala      50288  CTT GGG GGA GCC CCC TAC CAG GTG ACG GGC TGT GCC CTG GGT TTC TTC     33551  Leu Gly Gly Ala Pro Tyr Gln Val Thr Gly Cys Ala Leu Gly Phe Phe      66336  CTT CAG ATC TAC CTA TTG GAT GTG GCT CAG GTG GGC CCT TTC TCT GCC     38367  Leu Gln Ile Tyr Leu Leu Asp Val Ala Gln Val Gly Pro Phe Ser Ala      82384  TCC ATC ATC CTG TTT GTG GGC CGA GCC TGG GAT GCC ATC ACA GAC CCC     43183  Ser Ile Ile Leu Phe Val Gly Arg Ala Trp Asp Ala Ile Thr Asp Pro      98432  CTG GTG GGC CTC TGC ATC AGC AAA TCC CCC TGG ACC TGC CTG GGT CGC     47999  Leu Val Gly Leu Cys Ile Ser Lys Ser Pro Trp Thr Cys Leu Gly Arg     114480  CTT ATG CCC TGG ATC ATC TTC TCC ACG CCC CTG GCC GTC ATT GCC TAC     527115  Leu Met Pro Trp Ile Ile Phe Ser Thr Pro Leu Ala Val Ile Ala Tyr     130528  TTC CTC ATC TGG TTC GTG CCC GAC TTC CCA CAC GGC CAG ACC TAT TGG     575131  Phe Leu Ile Trp Phe Val Pro Asp Phe Pro His Gly Gln Thr Tyr Trp     146576  TAC CTG CTT TTC TAT TGC CTC TTT GAA ACA ATG GTC ACG TGT TTC CAT     623147  Tyr Leu Leu Phe Tyr Cys Leu Phe Glu Thr Met Val Thr Cys Phe His     162624  GTT CCC TAC TCG GCT CTC ACC ATG TTC ATC AGC ACC GAG CAG ACT GAG     671163  Val Pro Tyr Ser Ala Leu Thr Met Phe Ile Ser Thr Glu Gln Thr Glu     178672  CGG GAT TCT GCC ACC GCC TAT CGG ATG ACT GTG GAA GTG CTG GGC ACA     719179  Arg Asp Ser Ala Thr Ala Tyr Arg Met Thr Val Glu Val Leu Gly Thr     194720  GTG CTG GGC ACG GCG ATC CAG GGA CAA ATC GTG GGC CAA GCA GAC ACG     767195  Val Leu Gly Thr Ala Ile Gln Gly Gln Ile Val Gly Gln Ala Asp Thr     210 768 CCT TGT TGT TTC CAC GAC CTC AAT AGC TCT ACA GCT TCA CAA AGT GCC 815211 Pro CYS PHE GLN ASN SER Val Ala Val Ala 2268816 AC Cat ACAGCAGCAGCAG GAG GAG GAG GAG GAG GAG GAG GAG GAG TC TC -CAC TC TC -C -C -CTA's G's TAC     863227  Asn His Thr His Gly Thr Thr Ser His Arg Glu Thr Gln Lys Ala Tyr     242864  CTG CTG GCA GCG GGG GTC ATT GTC TGT ATC TAT ATA ATC TGT GCT GTC     911243  Leu Leu Ala Ala Gly Val Ile Val Cys Ile Tyr Ile Ile Cys Ala Val     258912  ATC CTG ATC CTG GGC GTG CGG GAG CAG AGA GAA CCC TAT GAA GCC CAG     959259  Ile Leu Ile Leu Gly Val Arg Glu Gln Arg Glu Pro Tyr Glu Ala Gln     274960  CAG TCT GAG CCA ATC GCC TAC TTC CGG GGC CTA CGG CTG GTC ATG AGC    1007275  Gln Ser Glu Pro Ile Ala Tyr Phe Arg Gly Leu Arg Leu Val Met Ser     2901008  CAC GGC CCA TAC ATC AAA CTT ATT ACT GGC TTC CTC TTC ACC TCC TTG    1055291  His Gly Pro Tyr Ile Lys Leu Ile Thr Gly Phe Leu Phe Thr Ser Leu     3061056  GCT TTC ATG CTG GTG GAG GGG AAC TTT GTC TTG TTT TGC ACC TAC ACC    1103307  Ala Phe Met Leu Val Glu Gly Asn Phe Val Leu Phe Cys Thr Tyr Thr     3221104  TTG GGC TTC CGC AAT GAA TTC CAG AAT CTA CTC CTG GCC ATC ATG CTC    1151323  Leu Gly Phe Arg Asn Glu Phe Gln Asn Leu Leu Leu Ala Ile Met Leu     3381152  TCG GCC ACT TTA ACC ATT CCC ATC TGG CAG TGG TTC TTG ACC CGG TTT    1199339  Ser Ala Thr Leu Thr Ile Pro Ile Trp Gln Trp Phe Leu Thr Arg Phe     3541200  GGC AAG AAG ACA GCT GTA TAT GTT GGG ATC TCA TCA GCA GTG CCA TTT    1247355  Gly Lys Lys Thr Ala Val Tyr Val Gly Ile Ser Ser Ala Val Pro Phe     3701248  CTC ATC TTG GTG GCC CTC ATG GAG AGT AAC CTC ATC ATT ACA TAT GCG    1295371  Leu Ile Leu Val Ala Leu Met Glu Ser Asn Leu Ile Ile Thr Tyr Ala     3861296  GTA GCT GTG GCA GCT GGC ATC AGT GTG GCA GCT GCC TTC TTA CTA CCC    1343387  Val Ala Val Ala Ala Gly Ile Ser Val Ala Ala Ala Phe Leu Leu Pro     4021344  TGG TCC ATG CTG CCT GAT GTC ATT GAC GAC TTC CAT CTG AAG CAG CCC    1391403  Trp Ser Met Leu Pro Asp Val Ile Asp Asp Phe His Leu Lys Gln Pro     4181392  CAC TTC CAT GGA ACC GAG CCC ATC TTC TTC TCC TTC TAT GTC TTC TTC    1439419  His Phe His Gly Thr Glu Pro Ile Phe Phe Ser Phe Tyr Val Phe Phe     4341440  ACC AAG TTT GCC TCT GGA GTG TCA CTG GGC ATT TCT ACC CTC AGT CTG    1487435  Thr Lys Phe Ala Ser Gly Val Ser Leu Gly Ile Ser Thr Leu Ser Leu     4501488  GAC TTT GCA GGG TAC CAG ACC CGT GGC TGC TCG CAG CCG GAA CGT GTC    1535451  Asp Phe Ala Gly Tyr Gln Thr Arg Gly Cys Ser Gln Pro Glu Arg Val     4661536  AAG TTT ACA CTG AAC ATG CTC GTG ACC ATG GCT CCC ATA GTT CTC ATC    1583467  Lys Phe Thr Leu Asn Met Leu Val Thr Met Ala Pro Ile Val Leu Ile     4821584  CTG CTG GGC CTG CTG CTC TTC AAA ATG TAC CCC ATT GAT GAG GAG AGG    1631 483 Leu Leu Gly Leu Leu Leu Phe Lys Met Tyr Pro Ile Asp Glu Glu Arg    4981632 CGG CGG CAG AAT AAG AAG GCC CTG CAG GCA CTG AGG GAC GAG GCC AGC   1679499 Arg Arg Gln Asn Lys Lys Ala Leu Gln Ala Leu Arg Asp Glu Ala Ser    5141680 AGC TCT GGC TGC TCA GAA ACA GAC TCC ACA GAG CTG GCT AGC ATC CTC   1727515 Ser Ser Gly Cys Ser Glu Thr Asp Ser Thr Glu Leu Ala Ser Ile Leu    5301728 TAG GGC CCG CCA CGT TGC CCG AAG CCA CCA TGC AGA AGG CCA CAG AAG   1775531 * **                                                                5311776 GGA TCA GGA CCT GTC TGC CGG CTT GCT GAG CAG CTG GAC TGC AGG TGC   18231824 TAG GAA GGG AAC TGA AGA CTC AAG GAG GTG GCC CAG GAC ACT TGC TGT   18711872 GCT CAC TGT GGG GCC GGC TGC TCT GTG GCC TCC TGC CTC CCC TCT GCC   19191920 TGC CTG TGG GGC CAA GCC CTG GGG CTG CCA CTG TGA ATA TGC CAA GGA   19671968 CTG ATC GGG CCT AGC CCG GAA CAC TAA TGT AGA AAC CTT TTT TTT ACA   20152016 GAG CCT AAT TAA TAA CTT AAT GAC TGT GTA CAT AGC AAT GTG TGT GTA 2062064 TGT ATA TGT CTG TGA GCT ATT AAT AAT AAT TTT CAT AAA AGC TGG 2112112 AAA GCA AAA AAA AAA AAA AAA AAA AAA AAA AAA Aaa

4.PP9445

A：核苷酸序列(SEQ ID NO：10)长度：1831bp1  GCCGCCGCGG  AGCGAGGTTG  ACTGGAGAGA  GCGCCTGGGC  GCAGAAGGGT  TAACGGGCCA61  CCGGGGGCTC  GCAGAGCAGG  AGGGTGCTCT  CGGACGGTGT  GTCCCCCACT  GCACTCCTGA121  ACTTGGAGGA  CAGGGTCGCC  GCGAGGGACG  CAGGTGGGTG  CCCTTGATCC  AGCTCAGCCC181  GATGGCAGAA  GAGGTTGACA  AAAAAGAAAG  ACACCTGTTG  GGGTGGCCTG  CCAGACCCAG241  GAGTGGAGGG  CTCTGTGAGG  GCCCGGGAAT  TCGGACTCAG  GACAGGGATT  CTCCATGGCT301  AGGCCCAGAA  ACACAGGGTC  CAACCACTCT  CCAGCAGGGA GACCTGGGGG  TGAAGGGGTG361  AGCCCTGCGC  AGGTCTCTGT  TCCTTGGTCT  TCACTGGGCA  GTGTGGAGAG  GTGTGGCCAG421  GAGGAGCCCG  CGTTTGTCCA  GAGCAGGGTC  TACTCTGGCA  CCAGAGTGAC  CACCTCTGAC481  CTCTCCTTTC  CTCGTCCTGG  GCCGGGAACG  ACACCAAATG  AGGGACATGG  AAAGGGCTGG541  AGTAACAAGA  GTCAGGCAGA  GCCTGAAGAC  TTGGGTGGAA  CATGGGCCCT  TCTCTGGAGA601  TCCTGGCCTC  CCCCGTTCAG  TCAGGGTGGA  GTTGCTGACC  TTAGTGGCCG  GCCCAGCCAG661  GGGAAGGAGT  GGCCATCGGC  AACCCCCACC  CCAACCCCAA  TCCCTGAGGC  GCCCGCTCTG721  GCTCAGCCAC  TCTGACCCCT  CCCTCAAATT  CCGAACCCTA  GGTCTCAGGG  AGGGCAGTGG781  GGCTGAGTGT  CTGCCCCCAG  GCACATTCCT  ACCCTTCTCT  TGGTCATTTT  CTGCCCCAGA841 GCTGGCCCAC  CTCAGCAATG  CGAGGGCTCC  CTGGATTCCT  CTCCCGGGTG  CCTTTCAGAT901  CCAACAGAAA  CAGATTTTTT  TTTTCCTGGA  AAGCAGAACT  AAGAGTGGGA  TGAGGAGCAG961  GGGTGGGAAG  GACTCAAAGT  GAGAAGAAGG  GGGCAAAGAG  AGTCAGGCTT  GGTGGCTGGG1021  GTGGCTTCCA  AGCCTCACTT  CTCCAGTGTT  CAAAGCTGAA  CTTCAGATGG  ACTTCCCGGC1081  TCTTCAGAAT  GAGAGGCCTG  TGGCTGGGGC  ATGAGGCAGC  CCCGGCTGCA  CCTCTCCTTC1141  CCGCTTCCCC  AGCTGGTAGA  GACGCACAGG  AAACAAGCCC  TCACTGAACC  AACTCCAGAT1201  GCTGGCACCC  AGAGTGGGTG  TTACATTGCC  GGCTTCTTCT  CTAGAGATTA  AACCGTCAAC1261  CCATTTAGCT  TATCCCTTGG  CCAAAAAGTG  TATGAGATGT  GCCTGGATGT  TCCCTAAAGA1321  GCTTATCTAA  GAAGGGAAGA GAAAGCCGGG  AGGCAAGTAG  GACAGAGAGA  TGACTGGGGA1381  AGGTCTTGTG  TCTGGAAGAC  CCAAGGAAGG  GGCTTCTGGT  GGGTCCTCAG  AGAGAGTGTC1441  TGGCGCATCC  TCAGTGGAGC  CTTCCTCCTC  TACTTTCTAG  GCACCTCTGG  GAGGGCAGGA1501  GTGGGAGCAG  ATGACAACCA  TTTTAGAAGG  AGCCCTCTGG  CTGGGTGCGG  TGGCTCACAC1561  CTGTCATCCC  AGCACTTTGG  GAGGCCAAGG  CAGGAGAAGC  GCTTGAGGCC  TGGAGTTCAA1621  GACCAGCCTG  TGCAATTTAG  CTGGATCCCA  TCTCCACCAA  AAAATACCAA  AATTAGCTGG1681  GTGTGGTGGT GCACGCATGT AGTCCCACCT ACTCAGGAGG CTGAGGAAGG AGAGCCTGTG1741  AGTTTGAGGC TGCAATGAGC TTTGGTGGCA CCACTGCCCT CCAGCCTGGA TGACAGAGTG1801  AGATCTCCAT CTCAAAAAAA AAAAAAAAAA A

B: Amino acid sequence (SEQ ID NO: 11) length: 154 amino acids

C. Nucleotide and amino acid combination sequence (SEQ ID NO: 12)

Clone number: PP9445

Start code: 518 ATG End code: 980 TGA

蛋白质分子量：17150.751    G CCG CCG CGG AGC GAG GTT GAC TGG AGA GAG CGC CTG GGC GCA GAA      4647  GGG TTA ACG GGC CAC CGG GGG CTC GCA GAG CAG GAG GGT GCT CTC GGA      9495  CGG TGT GTC CCC CAC TGC ACT CCT GAA CTT GGA GGA CAG GGT CGC CGC     142143  GAG GGA CGC AGG TGG GTG CCC TTG ATC CAG CTC AGC CCG ATG GCA GAA     190191  GAG GTT GAC AAA AAA GAA AGA CAC CTG TTG GGG TGG CCT GCC AGA CCC     238239  AGG AGT GGA GGG CTC TGT GAG GGC CCG GGA ATT CGG ACT CAG GAC AGG     286287  GAT TCT CCA TGG CTA GGC CCA GAA ACA CAG GGT CCA ACC ACT CTC CAG     334335  CAG GGA GAC CTG GGG GTG AAG GGG TGA GCC CTG CGC AGG TCT CTG TTC     382383  CTT GGT CTT CAC TGG GCA GTG TGG AGA GGT GTG GCC AGG AGG AGC CCG     430431  CGT TTG TCC AGA CCA GGG TCT ACT CTG GCA CCA GAG TGA CCA CCT CTG     478479  ACC TCT CCT TTC CTC GTC CTG GGC CGG GAA CGA CAC CAA ATG AGG GAC     5261                                                      Met Arg Asp       3527  ATG GAA AGG GCT GGA GTA ACA AGA GTC AGG CAG AGC CTG AAG ACT TGG     5744  Met Glu Arg Ala Gly Val Thr Arg Val Arg Gln Ser Leu Lys Thr Trp      19575  GTG GAA CAT GGG CCC TTC TCT GGA GAT CCT GGC CTC CCC CGT TCA GTC     62220  Val Glu His Gly Pro Phe Ser Gly Asp Pro Gly Leu Pro Arg Ser Val      35623  AGG GTG GAG TTG CTG ACC TTA GTG GCC GGC CCA GCC AGG GGA AGG AGT     67036  Arg Val Glu Leu Leu Thr Leu Val Ala Gly Pro Ala Arg Gly Arg Ser      51671  GGC CAT CGG CAA CCC CCA CCC CAA CCC CAA TCC CTG AGG CGC CCG CTC     71852  Gly His Arg Gln Pro Pro Pro Gln Pro Gln Ser Leu Arg Arg Pro Leu      67719  TGG CTC AGC CAC TCT GAC CCC TCC CTC AAA TTC CGA ACC CTA GGT CTC     76668  Trp Leu Ser His Ser Asp Pro Ser Leu Lys Phe Arg Thr Leu Gly Leu      83767  AGG GAG GGC AGT GGG GCT GAG TGT CTG CCC CCA GGC ACA TTC CTA CCC     81484  Arg Glu Gly Ser Gly Ala Glu Cys Leu Pro Pro Gly Thr Phe Leu Pro      99815  TTC TCT TGG TCA TTT TCT GCC CCA GAG CTG GCC CAC CTC AGC AAT GCG     862100  Phe Ser Trp Ser Phe Ser Ala Pro Glu Leu Ala His Leu Ser Asn Ala     115863  AGG GCT CCC TGG ATT CCT CTC CCG GGT GCC TTT CAG ATC CAA CAG AAA     910116 ARG ALA Pro TRP I Pro Leu Pro Gly Ala Phe Gln Ile Gln GLN LYS 131 911 CAG ATT TTT TTC CTG GAA AGC AGT GGG AGG AGC AGC 958132 Gl Merile PHE Leu sele Ser     147959  AGG GGT GGG AAG GAC TCA AAG TGA GAA GAA GGG GGC AAA GAG AGT CAG    1006148  Arg Gly Gly Lys Asp Ser Lys ***                                     1551007  GCT TGG TGG CTG GGG TGG CTT CCA AGC CTC ACT TCT CCA GTG TTC AAA    10541055  GCT GAA CTT CAG ATG GAC TTC CCG GCT CTT CAG AAT GAG AGG CCT GTG    11021103  GCT GGG GCA TGA GGC AGC CCC GGC TGC ACC TCT CCT TCC CGC TTC CCC    11501151  AGC TGG TAG AGA CGC ACA GGA AAC AAG CCC TCA CTG AAC CAA CTC CAG    11981199  ATG CTG GCA CCC AGA GTG GGT GTT ACA TTG CCG GCT TCT TCT CTA GAG    12461247  ATT AAA CCG TCA ACC CAT TTA GCT TAT CCC TTG GCC AAA AAG TGT ATG    12941295  AGA TGT GCC TGG ATG TTC CCT AAA GAG CTT ATC TAA GAA GGG AAG AGA    13421343  AAG CCG GGA GGC AAG TAG GAC AGA GAG ATG ACT GGG GAA GGT CTT GTG    13901391  TCT GGA AGA CCC AAG GAA GGG GCT TCT GGT GGG TCC TCA GAG AGA GTG    14381439  TCT GGC GCA TCC TCA GTG GAG CCT TCC TCC TCT ACT TTC TAG GCA CCT    14861487 CTG GGA GGG CAG GAG TGG GAG CAG ATG ACA ACC ATT TTA GAA GGA GCC    15341535  CTC TGG CTG GGT GCG GTG GCT CAC ACC TGT CAT CCC AGC ACT TTG GGA    15821583  GGC CAA GGC AGG AGA AGC GCT TGA GGC CTG GAG TTC AAG ACC AGC CTG 16301631  TGC AAT TTA GCT GGA TCC CAT CTC CAC CAA AAA ATA CCA AAA TTA GCT    16781679  GGG TGT GGT GGT GCA CGC ATG TAG TCC CAC CTA CTC AGG AGG CTG AGG    17261727  AAG GAG AGC CTG TGA GTT TGA GGC TGC AAT GAG CTT TGG TGG CAC CAC 17741775 TGC CCT CCA GCC TGG ATG ACA GAG TGA GAT CTC CAT CTC AAA AAA AAA 18221823 AAA AAA AAA 1 8 8

5. PP10199

A：核苷酸序列(SEQ ID NO：13)长度：1739bp1    GTTCCAGAGC  CACTTTTAAG  ATTCTTCAAT  TCCAAATGCA  TGTCTTTTTT  TAAAAAAAAG61    AAAGAAAGAA  AAATAAGTTT  CTAATATTAG  AGAAGTACAG  CCCTGAATTG  GGTTTTGTGT121    CCACTGCTGG  ACCCCATGAG  GGCCAGGTGG  AGTGGACCTC  TGCAGCCCCA  GTTGTGTGCA181    CTCTCTGTTT  GGTGCAAATT  CCAGTTTGCT  GGTTCTCAAT  AGCAAGACCA  GCCTGAGACC241    ACCTGTCCTG  CTCTTCCCAT  GAGAGGGCCG  AATGCTCCCA  GCCTCCATGC  CATGTCCTGT301    TCCTGGGGTC  CTGGGGGTCA  TTGCAGCCTG  TATGTGCTTC CTCCAGCCAG  GGTGATCATC361    GGGTGCCCCA  GTGAGCCCCA  GCACTGAGGG  TCAGCCCCAG  GCACTGTCAA  AGGTGAGAGC421    TCAGAGGCTG  TGCCCAGAAA  GAGAGGTGGG  CCCTGCCTGC  CCTGGACGGA  GGGAGAGAGG481    CTTCTCAGAG  CCCGAGGCAT  GAACCCTCAG  GTGGGTCGTG  GCCATAGTCA  GATGATGGCT541    GCTGGTGAGC  TCAGTGACCA  GGCGTCTTCA  GGCAGCTCAT  AAGTTTGAGA  GGACACAGCC601    TAAGGGAGGT  TTGCTGGGGA  GTAGCCCCAC  TTCCACCCTG  AATAGACAAG  AGATGGTAAA661    GCAGGTACCC  AGCACTTAGT  GCTTTCTTGG  GGATATCGCG  TGGGTCCCCG  GGGGCCTGGG721    TGCCCGAAGT  GCCGCAGTAC  TCCATGGTGC  AGAGAGCTTG  CTCCTGTGGA  GGAAGTGTCT781    ATGTGGTCCC  CAGCTCCTCT  GTCTGCCTGT  CCACTGAGGG  GCACCCATGG  CTCAGCAGAA841 GGGCTATTCT  TGGGGTTCCC  GGTCCTCCTC  CAGCCCCGCT  AATCTGTGTA  GGCCTCAAGT901    GCTGTGTGTT  TGTAAGCATT  GTCATCCACA  GTCCTATTGT  ACGAGCTGGT  TCACCCGCAG961    CTCTGAGCTG  CTCTCCAGCC  CCAGCCCTTT  CTTCCTGTGC  CCCTACCCCC  GCTGGGATGA1021    CTCTCCTCAC  CCTCCCTGGG  GCGACAACCG  CCCTGTCTGT  AATGAGTGGC  AGTCCCAAGC1081    TTCCTGACTG  GCTTCCGCAG  CTCTCTGACT  CCCCTAAACA  AGGCCTCAGG  GACTCCACAT1141    CCAAATTAAG  GCGGCACCTG  GTGGCAGGTT  GGCATTTTCC  GGTGTCCTAT  CTATGAAAGA1201    CAGGAAGACA  GCTGGGAGCA  AACTCCCCTG  GGCCAGACTC  TTGGAAACAT  AAAGGCTTGG1261    GTGCCCAGCT  GGGGACCGGG  AGAAAGTCTA  AAACACGGGA  CTGGGCCAAG  GACCCCACAG1321    GTCCCTGTCT  CATTAGGTCC CCTGAAACGT  GTGGAAGCTA  AAATGGCATT  CACGTGATTC1381    TTGATCATTT  AACAGTGGAT  TCTGATCTGA  TACTACACTG  AGAAGTGCCC  CTGGGCCGGG1441  CGCGGTGGCT CACGCCTGTA ATCCCAGCAC TTTGGGAGGC CGAGGCGGGC GGATCACAAG1501  GAGATTGAGA CCATCCTGGC TAACACGGTG AAACCCTGTC TCTACTAAAA ATACAAAAAA1561  TTAGCCAGGC ATGGTGGCAG GCGCCTCTAG TACTAGCTAC TCGGGAGGCT GAGGCAGGAG1621  AATGGTGTGA ACCCGGGAGG CGGAACTTGC AGTGAGCCAA GATTGTGCCA CTGCACTCTA1681  GCATGGGCGA CAGAGCAAGA CTCAGTCTCA AAAAAAAAAA AAAAAAAAAA AAAAAAAAA

B: amino acid sequence (SEQ ID NO: 14) length: 150 amino acids

C. Nucleotide and amino acid combination sequence (SEQ ID NO: 15)

Clone number: PP10199

Start code: 744 ATG End code: 1194 TGA

蛋白质分子量：15496.341   GT TCC AGA GCC ACT TTT AAG ATT CTT CAA TTC CAA ATG CAT GTC TTT      4748  TTT TAA AAA AAA GAA AGA AAG AAA AAT AAG TTT CTA ATA TTA GAG AAG      9596  TAC AGC CCT GAA TTG GGT TTT GTG TCC ACT GCT GGA CCC CAT GAG GGC     143144  CAG GTG GAG TGG ACC TCT GCA GCC CCA GTT GTG TGC ACT CTC TGT TTG     191192  GTG CAA ATT CCA GTT TGC TGG TTC TCA ATA GCA AGA CCA GCC TGA GAC     239240  CAC CTG TCC TGC TCT TCC CAT GAG AGG GCC GAA TGC TCC CAG CCT CCA     287288  TGC CAT GTC CTG TTC CTG GGG TCC TGG GGG TCA TTG CAG CCT GTA TGT     335336  GCT TCC TCC AGC CAG GGT GAT CAT CGG GTG CCC CAG TGA GCC CCA GCA     383384  CTG AGG GTC AGC CCC AGG CAC TGT CAA AGG TGA GAG CTC AGA GGC TGT     431432  GCC CAG AAA GAG AGG TGG GCC CTG CCT GCC CTG GAC GGA GGG AGA GAG     479480  GCT TCT CAG AGC CCG AGG CAT GAA CCC TCA GGT GGG TCG TGG CCA TAG     527528  TCA GAT GAT GGC TGC TGG TGA GCT CAG TGA CCA GGC GTC TTC AGG CAG     575576  CTC ATA AGT TTG AGA GGA CAC AGC CTA AGG GAG GTT TGC TGG GGA GTA     623624  GCC CCA CTT CCA CCC TGA ATA GAC AAG AGA TGG TAA AGC AGG TAC CCA     671672  GCA CTT AGT GCT TTC TTG GGG ATA TCG CGT GGG TCC CCG GGG GCC TGG     719720  GTG CCC GAA GTG CCG CAG TAC TCC ATG GTG CAG AGA GCT TGC TCC TGT     7671                                  Met Val Gln Arg Ala Cys Ser Cys       8768  GGA GGA AGT GTC TAT GTG GTC CCC AGC TCC TCT GTC TGC CTG TCC ACT 8159 Gly Gly Ser Val Tyr Val Val Pro Ser Seer Val Cys Leu Ser 24816 GGG CAC CAC CCA TGG CTC AGGC AGG GCT CTT GGG GGG GGT 8632 GLULY ARGLRL's ARGRL's HRT -GTC's C's GTC -C's Pro Gly      40864  CCT CCT CCA GCC CCG CTA ATC TGT GTA GGC CTC AAG TGC TGT GTG TTT     91141  Pro Pro Pro Ala Pro Leu Ile Cys Val Gly Leu Lys Cys Cys Val Phe      56912  GTA AGC ATT GTC ATC CAC AGT CCT ATT GTA CGA GCT GGT TCA CCC GCA     95957  Val Ser Ile Val Ile His Ser Pro Ile Val Arg Ala Gly Ser Pro Ala      72960  GCT CTG AGC TGC TCT CCA GCC CCA GCC CTT TCT TCC TGT GCC CCT ACC    100773  Ala Leu Ser Cys Ser Pro Ala Pro Ala Leu Ser Ser Cys Ala Pro Thr      881008  CCC GCT GGG ATG ACT CTC CTC ACC CTC CCT GGG GCG ACA ACC GCC CTG    105589  Pro Ala Gly Met Thr Leu Leu Thr Leu Pro Gly Ala Thr Thr Ala Leu     1041056  TCT GTA ATG AGT GGC AGT CCC AAG CTT CCT GAC TGG CTT CCG CAG CTC    1103105  Ser Val Met Ser Gly Ser Pro Lys Leu Pro Asp Trp Leu Pro Gln Leu     1201104  TCT GAC TCC CCT AAA CAA GGC CTC AGG GAC TCC ACA TCC AAA TTA AGG    1151121  Ser Asp Ser Pro Lys Gln Gly Leu Arg Asp Sering Thr Lys Leu ARG 1361152 CGG CAC CTG GCA GCA GGT TGG Cat TTT CCG GTG TAT CTA TGA AAG 1199137 ARG His Leu Val Ala Gly Pro Valr Leu *** 1511200 ACA GAGA AC TCC CCT GGG CCA GAC TCT TGG AAA    12471248  CAT AAA GGC TTG GGT GCC CAG CTG GGG ACC GGG AGA AAG TCT AAA ACA    12951296  CGG GAC TGG GCC AAG GAC CCC ACA GGT CCC TGT CTC ATT AGG TCC CCT    13431344  GAA ACG TGT GGA AGC TAA AAT GGC ATT CAC GTG ATT CTT GAT CAT TTA    13911392  ACA GTG GAT TCT GAT CTG ATA CTA CAC TGA GAA GTG CCC CTG GGC CGG    14391440  GCG CGG TGG CTC ACG CCT GTA ATC CCA GCA CTT TGG GAG GCC GAG GCG    14871488  GGC GGA TCA CAA GGA GAT TGA GAC CAT CCT GGC TAA CAC GGT GAA ACC    15351536  CTG TCT CTA CTA AAA ATA CAA AAA ATT AGC CAG GCA TGG TGG CAG GCG    15831584  CCT CTA GTA CTA GCT ACT CGG GAG GCT GAG GCA GGA GAA TGG TGT GAA    16311632  CCC GGG AGG CGG AAC TTG CAG TGA GCC Aag Att GTG CCA CTG CAC TCT 16791680 AGC ATG GGC GAC AGA GCA AGA CTC AGT CTC AAA AAA AAA AAA 17271728 AAA AAA 17399

6. PP10226

A：核苷酸序列(SEQ ID NO：16)长度：1012bp1  GTGAGAGAGG  GGTTTGGAAA  TACCAGACTA  TAATTGTGGA  TTTGTCCATT  ACTCCTTTCA61  GTTCTAGCAG  TTTTTGCTTC  TTGTGTTTTG  AAGCTCTGTT  ATTTGATAAA  AATTTTTAGA121  ATTTTTAATG  TTTATTTTAG  AATGTATAAA  ATTTTAGAAT  TTATATGGAT  AAATTGAATC181  CTCTATCATT  ATAACATTAT  GTTCTTTATG  CCTGTAATAT  TTTTTGCTGC  AAAATCTACT241  GTCTTAAATA  ATATAGACAC  AACAGCCTGA  TTAGTGTTTG  CATAGTACAT  CTTCCCCTTC301  TTCCATTGTT  TTACATTTAG  CCTATTTGTG  CTTTAAAAAA ATTTAAGTAC  CTATATTGTA361  GGCAGCATAG  AGTTGGATCT  TGTTTTATTA  ATGCACCCTG  TTTTGAGAGA  GAGAGAGAGA421  GAGACAGAGA  CAGAGACACA  GAGAGAGAGT  GTGAGCGAGC  AAAAGAGATT  TATTCTGGTT481  TTTTTTTGTT  TGTTTTTGAG  ATGGAGTCTT  GCTCTCTTGC  TCAGGCTGGA  GTGCAGTGCC541  GCAATCTCAG  CTCACTGCAA  CCTCCACCTC  CTGGGTTCAA  GTTATTCTCC  TGTCTCAGCC601  TCCCAAGTAG  CTGGGACTAC  AGGCCTGTGC  CACCATGCCC  GGCTACGTTT  TGTATTTTTA661  GTACAGACGG  TGTTTCACCA  TGTTGGCCAG  CCTGGTCTCA  AACTCCTGGC  CTCAAGTTGA721  TCTGCTGGCC  TCACGCCTGT  AATCCTAGTA  CTTTGGGAGG  CCGAGGCGGG  CGGATCTCGA781  GTTCAGGAGA  TCGACCATCC  TGGCTAACAC  GGTGAAACCT  CGTCTCTACT  AAAAATACAA841 AAAATTAGCC GGGCATGGTG GTGGGCACCC GTAGTCCCAG CTACTTGGGA GGCTGAGGCA901 GGAGAATGGC ATGAATCCAG TAGGCGGAGC TTGCAGTGAG CCAAGATCAC GCCACTGCAC961 TCCACCCTGG GTGAAAGAGC GA GACTTTGT AAAAAAAA

B: amino acid sequence (SEQ ID NG: 17) length: 109 amino acids

C. Nucleotide and amino acid combination sequence (SEQ ID NO: 18)

Clone No.: PP10226 Start Codon: 391 ATG Stop Codon: 718 TGA Protein Molecular Weight: 12262.591 GTG AGA GAG GGG TTT GGA AAT ACC AGA CTA TAA TTG TGG ATT TGT CCA CT T 4849 G GTT CTC CTT TCA GT CT GTT TTG AAG CTC 997 TAT TAA AAA AAA AAA AGA Attt TAT GTT TAT TAGA AGA AGA AGA AGA AGA AGA Atg 144145 TAAA ATT TTA TGG ATA ATC CTC AAC Atg TTC TTT AAC Atg TTT A GCT GCA AAA TCT ACT     240241  GTC TTA AAT AAT ATA GAC ACA ACA GCC TGA TTA GTG TTT GCA TAG TAC     288289  ATC TTC CCC TTC TTC CAT TGT TTT ACA TTT AGC CTA TTT GTG CTT TAA     336337  AAA AAT TTA AGT ACC TAT ATT GTA GGC AGC ATA GAG TTG GAT CTT GTT     384385  TTA TTA ATG CAC CCT GTT TTG AGA GAG AGA GAG AGA GAG ACA GAG ACA     4321          Met His Pro Val Leu Arg Glu Arg Glu Arg Glu Thr Glu Thr      14433  GAG ACA CAG AGA GAG AGT GTG AGC GAG CAA AAG AGA TTT ATT CTG GTT     48015  Glu Thr Gln Arg Glu Ser Val Ser Glu Gln Lys Arg Phe Ile Leu Val      30481  TTT TTT TGT TTG TTT TTG AGA TGG AGT CTT GCT CTC TTG CTC AGG CTG     52831  Phe Phe Cys Leu Phe Leu Arg Trp Ser Leu Ala Leu Leu Leu Arg Leu      46529  GAG TGC AGT GGC GCA ATC TCA GCT CAC TGC AAC CTC CAC CTC CTG GGT     57647  Glu Cys Ser Gly Ala Ile Ser Ala His Cys Asn Leu His Leu Leu Gly      62577  TCA AGT TAT TCT CCT GTC TCA GCC TCC CAA GTA GCT GGG ACT ACA GGC     62463  Ser Ser Tyr Ser Pro Val Ser Ala Ser Gln Val Ala Gly Thr Thr Gly      78625  CTG TGC CAC CAT GCC CGG CTA CGT TTT GTA TTT TTA GTA CAG ACG GTG     67279  Leu Cys His His Ala Arg Leu Arg PHTTT, PHTTT, PHR PHTTT, PHR PHR PHR GLN TTT GGT GGT, CAC, CTC GCC, has His His His Leu Leu Ala, Gly Val His His Leu Ala? CCT GTA ATC CTA GTA CTT TGG GAG GCC GAG GCG     768769  GGC GGA TCT CGA GTT CAG GAG ATC GAC CAT CCT GGC TAA CAC GGT GAA     816817  ACC TCG TCT CTA CTA AAA ATA CAA AAA ATT AGC CGG GCA TGG TGG TGG     864865  GCA CCC GTA GTC CCA GCT ACT TGG GAG GCT GAG GCA GGA GAA TGG CAT     912913  GAA TCC AGT AGG CGG AGC TTG CAG TGA GCC AAG ATC ACG CCA CTG CAC     960961  TCC AGC CTG GGT GAC AGA GCG AGA CTT TGT CTC AAA AAA AAA AAA AAA    10081009  AAA A                                                              1012

7. SP2114a

A：核苷酸序列(SEQ ID NO：19)长度：2546bp1  GGCCAGTCAA  GATGGCCGCC  GCTGGGTGAG  GCAAGCTGGC  GCGCCGCGGG  GGCGTCTGGG61  AGTTGTAGTT  CGGGACGGCG  GGCTGACGCA  CTTCGCCGCC  GGCCGACGGG  CGCCATTGTG121  CGGCGCGCGC  CGGGACTCTG  CCCACTTCCA  CCAGAGACAC  ATTGAGAAGG  AGGAAACTAT181  GGCCTCCAGG  CTTCCGACGG  CCTGGTCCTG  TGAACCAGAG  ACCTTTGAAG  ATCTAACACT241  GGGTTTTACC  CCGGAAGAGT  GGGGACTGCT  GGACCTCAAA  CAGAAGTCCC  TGTACAGGGA301  AGTGATGCTG  GAGAACTACA  GGAACCTGGT  CTCAGTGGAA CATCAGCTTT  CCAAACCAGA361  TGTGGTATCT  CAGTTAGAGG  AGGCAGAAGA  TTTCTGGCCA  GTGGAGAGAG  GAATTCCTCA421  AGACACCATT  CCTGAGTATC  CTGAGCTCCA  GCTGGACCCT  AAATTGGATC  CTCTTCCTGC481  TGAGAGTCCC  CTAATGAACA  TTGAGGTTGT  TGAGGTCCTC  ACACTGAACC  AGGAGGTGGC 541    TGGTCCCCGG  AATGCCCAGA  TCCAGGCCCT  ATATGCTGAA  GATGGAAGCC  TGAGTGCAGA601    TGCCCCCAGT  GAGCAGATCC  AACAGCAGGG  CAAGCATCCA  GGTGACCCTG  AGGCCGCGCG661    CCAGAGGTTC  CGGCAGTTCC  GTTATAAGGA  CATGACAGGT  CCCCGGGAGG  CCCTGGACCA721    GCTCCGAGAG  CTGTGTCACC  AGTGGCTACA  GCCTAAGGCA  CGCTCCAAGG  AGCAGATCCT781    GGAGCTGCTG  GTGCTGGAGC  AGTTCCTAGG  TACACTGCCT  GTGAAGCTCC GGACATGGGT841    GGAATCGCAG  CACCCAGAGA  ACTGCCAAGA  GGTGGTGGCC  CTGGTAGAGG  GTGTGACCTG901    GATGTCTGAG  GAGGAAGTAC  TTCCTGCAGG  ACAACCTGCC  GAGGGCACCA  CCTGCTGCCT961    CGAGGTCACT  GCCCAGCAGG  AGGAGAAGCA  GGAGGATGCA  GCCATCTGCC  CAGTGACAGT1021    GCTCCCTGAG  GAGCCAGTGA  CCTTCCAGGA  TGTGGCTGTG  GACTTCAGCC  GGGAGGAGTG1081    GGGGCTGCTG  GGCCCGACAC  AGAGGACCGA  GTACCGCGAT  GTGATGCTGG  AGACCTTTGG1141    GCACCTGGTC  TCTGTGGGGT  GGGAGACTAC  ACTGGAAAAT  AAAGAGTTAG  CTCCAAATTC1201    TGACATTCCT  GAGGAAGAAC  CAGCCCCCAG  CCTGAAAGTA  CAAGAATCCT  CAAGGGATTG1261    TGCCTTGTCC  TCTACATTAG  AAGATACCTT  GCAGGGTGGG  GTCCAGGAAG  TCCAAGACAC1321    AGTGTTGAAG CAGATGGAGT  CTGCTCAGGA  AAAAGACCTT  CCTCAGAAGA  AGCACTTTGA1381    CAACCGTGAG  TCCCAGGCAA  ACAGTGGTGC  TCTTGACACA  AACCAAGTTT  CGCTCCAGAA1441    AATTGACAAC  CCTGAGTCCC  AGGCAAACAG  TGGCGCTCTT  GACACAAACC  AAGTTTTGCT1501    CCACAAAATT  CCTCCTAGAA  AACGATTGCG  CAAACGTGAC  TCACAAGTTA  AAAGTATGAA1561    ACATAATTCA  CGTGTAAAAA  TTCATCAGAA  GAGCTGTGAA  AGGCAAAAGG  CCAAGGAAGG1621    CAATGGTTGT  AGGAAAACCT  TCAGTCGGAG  TACTAAACAG  ATTACGTTTA  TAAGAATTCA1681    CAAGGGGAGC  CAAGTTTGCC  GATGCAGTGA  ATGTGGTAAA  ATATTCCGGA  ACCCAAGATA1741    CTTTTCTGTG  CATAAGAAAA  TCCATACCGG  AGAGAGGCCC  TATGTGTGTC  AAGACTGTGG1801    GAAAGGATTT  GTTCAGAGCT  CTTCCCTCAC ACAGCATCAG  AGAGTTCATT  CTGGAGAGAG1861    ACCATTTGAA  TGTCAGGAGT  GTGGGAGGAC  CTTCAATGAT  CGCTCAGCCA  TCTCCCAGCA1921    CCTGAGGACT  CACACTGGCG  CTAAGCCCTA  CAAGTGTCAG  GACTGTGGAA  AAGCCTTCCG1981    CCAGAGTTCC  CACCTCATCA  GACATCAGAG  GACTCACACC  GGGGAGCGCC  CATATGCATG2041    CAACAAATGT  GGAAAGGCCT  TCACCCAGAG  CTCACACCTT  ATTGGGCACC  AGAGAACCCA2101    CAATAGGACA  AAGCGAAAGA  AGAAACAGCC  TACCTCATAG  CTCTCAAGCC  AGTTGAAGAA2161    ACCTTGCCTT  TTCAGCTTGA  CCCTGCAATA  TAACATGCAC  AGGCCTGCTT  GTGAATCAGG2221    ACTGAATGTG  AAAGGGAAGT  ATTGAGTGAG  GACATTCCCA  AAACCAAAGG  ACAACTGAGG2281    AGACTGCCCA  GCACATAATG  AATAAATAAG  AAAATGAGTG  AGGAGTTATT AACATCATTT2341    GGAAAAAAGA  TTTCCCATTC  ACTTGATATT  GTTTGTTCAC  TCATTTAGTC  ATTAAAAGTG2401    AGATTAATAA  AATCTGAAAA  TGTTATATAA  TAACTTTAAA  AAGCCAGGTA  ATTAATAATC2461    TGCACTGATA  TTACATCCAC  AGTACCACAG  TATTTATGTG  TATGAATTAA  GGATTAAAAG2521    ATAATGTGGA  TAAAAAAAAA  AAAAAA

B：氨基酸序列(SEQ ID NO：20)      长度：653个氨基酸1    MASRLPTAWS  CEPETFEDVT  LGFTPEEWGL  LDLKQKSLYR  EVMLENYRNL  VSVEHQLSKP61    DVVSQLEEAE  DFWPVERGIP  QDTIPEYPEL  QLDPKLDPLP  AESPLMNIEV  VEVLTLNQEV121    AGPRNAQIQA  LYAEDGSLSA  DAPSEQIQQQ  GKHPGDPEAA  RQRFRQFRYK  DMTGPREALD181    QLRELCHQWL  QPKARSKEQI  LELLVLEQFL  GTLPVKLRTW  VESQHPENCQ  EVVALVEGVT241    WMSEEEVLPA  GQPAEGTTCC  LEVTAQQEEK  QEDAAICPVT  VLPEEPVTFQ  DVAVDFSREE301    WGLLGPTQRT  EYRDVMLETF  GHLVSVGWET TLENKELAPN  SDIPEEEPAP  SLKVQESSRD361    CALSSTLEDT  LQGGVQEVQD  TVLKQMESAQ  EKDLPQKKHF  DNRESQANSG  ALDTNQVSLQ421    KIDNPESQAN  SGALDTNQVL  LHKIPPRKRL  RKRDSQVKSM  KHNSRVKIHQ  KSCERQKAKE481    GNGCRKTFSR  STKQITFIRI  HKGSQVCRCS  ECGKIFRNPR  YFSVHKKIHT  GERPYVCQDC541    GKGFVQSSSL  TQHQRVHSGE  RPFECQECGR  TFNDRSAISQ  HLRTHTGAKP  YKCQDCGKAF601    RQSSHLIRHQ  RTHTGERPYA  CNKCGKAFTQ  SSHLIGHQRT  HNRTKRKKKQ  PTS

C. Nucleotide and amino acid combination sequence (SEQ ID NO: 21)

Clone number: SP2114a

Start code: 179 ATG Stop code: 2138 TAG

蛋白质分子量：74247.901    G GCC AGT CAA GAT GGC CGC CGC TGG GTG AGG CAA GCT GGC GCG CCG      4647  CGG GGG CGT CTG GGA GTT GTA GTT CGG GAC GGC GGG CTG ACG CAG TTC      9495  GCC GCC GGC CGA CGG GCG CCA TTG TGC GGC GCG CGC CGG GAC TCT GCC     142143  CAC TTC CAC CAG AGA CAC ATT GAG AAG GAG GAA ACT ATG GCC TCC AGG     1901                                                  Met Ala Ser Arg       4191  CTT CCG ACG GCC TGG TCC TGT GAA CCA GAG ACC TTT GAA GAT GTA ACA     2385  Leu Pro Thr Ala Trp Ser Cys Glu Pro Glu Thr Phe Glu Asp Val Thr      20239  CTG GGT TTT ACC CCG GAA GAG TGG GGA CTG CTG GAC CTC AAA CAG AAG     28621  Leu Gly Phe Thr Pro Glu Glu Trp Gly Leu Leu Asp Leu Lys Gln Lys      36287  TCC CTG TAC AGG GAA GTG ATG CTG GAG AAC TAC AGG AAC CTG GTC TCA     33437  Ser Leu Tyr Arg Glu Val Met Leu Glu Asn Tyr Arg Asn Leu Val Ser      52335  GTG GAA CAT CAG CTT TCC AAA CCA GAT GTG GTA TCT CAG TTA GAG GAG     38253  Val Glu His Gln Leu Ser Lys Pro Asp Val Val Ser Gln Leu Glu Glu      68383  GCA GAA GAT TTC TGG CCA GTG GAG AGA GGA ATT CCT CAA GAC ACC ATT     43069  Ala Glu Asp Phe Trp Pro Val Glu Arg Gly Ile Pro Gln Asp Thr Ile      84431  CCT GAG TAT CCT GAG CTC CAG CTG GAC CCT AAA TTG GAT CCT CTT CCT     47885  Pro Glu Tyr Pro Glu Leu Gln Leu Asp Pro Lys Leu Asp Pro Leu Pro     100479  GCT GAG AGT CCC CTA ATG AAC ATT GAG GTT GTT GAG GTC CTC ACA CTG     526101  Ala Glu Ser Pro Leu Met Asn Ile Glu Val Val Glu Val Leu Thr Leu     116527  AAC CAG GAG GTG GCT GGT CCC CGG AAT GCC CAG ATC CAG GCC CTA TAT     574117  Asn Gln Glu Val Ala Gly Pro Arg Asn Ala Gln Ile Gln Ala Leu Tyr     132575  GCT GAA GAT GGA AGC CTG AGT GCA GAT GCC CCC AGT GAG CAG ATC CAA     622133  Ala Glu Asp Gly Ser Leu Ser Ala Asp Ala Pro Ser Glu Gln Ile Gln     148623  CAG CAG GGC AAG CAT CCA GGT GAC CCT GAG GCC GCG CGC CAG AGG TTC     670149 Gln Gln Gly Lys His Pro Gly Asp Pro Glu Ala Ala Arg Gln Arg Phe     164671  CGG CAG TTC CGT TAT AAG GAC ATG ACA GGT CCC CGG GAG GCC CTG GAC     718165  Arg Gln Phe Arg Tyr Lys Asp Met Thr Gly Pro Arg Glu Ala Leu Asp 180719  CAG CTC CGA GAG CTG TGT CAC CAG TGG CTA CAG CCT AAG GCA CGC TCC     766181  Gln Leu Arg Glu Leu Cys His Gln Trp Leu Gln Pro Lys Ala Arg Ser     196767  AAG GAG CAG ATC CTG GAG CTG CTG GTG CTG GAG CAG TTC CTA GGT ACA     814197  Lys Glu Gln Ile Leu Glu Leu Leu Val Leu Glu Gln Phe Leu Gly Thr     212815  CTG CCT GTG AAG CTC CGG ACA TGG GTG GAA TCG CAG CAC CCA GAG AAC     862213  Leu Pro Val Lys Leu Arg Thr Trp Val Glu Ser Gln His Pro Glu Asn     228863  TGC CAA GAG GTG GTG GCC CTG GTA GAG GGT GTG ACC TGG ATG TCT GAG     910 229  Cys Gln Glu Val Val Ala Leu Val Glu Gly Val Thr Trp Met Ser Glu     244911  GAG GAA GTA CTT CCT GCA GGA CAA CCT GCC GAG GGC ACC ACC TGC TGC     958245  Glu Glu Val Leu Pro Ala Gly Gln Pro Ala Glu Gly Thr Thr Cys Cys     260959  CTC GAG GTC ACT GCC CAG CAG GAG GAG AAG CAG GAG GAT GCA GCC ATC    1006261  Leu Glu Val Thr Ala Gln Gln Glu Glu Lys Gln Glu Asp Ala Ala Ile     2761007  TGC CCA GTG ACA GTG CTC CCT GAG GAG CCA GTG ACC TTC CAG GAT GTG    1054277  Cys Pro Val Thr Val Leu Pro Glu Glu Pro Val Thr Phe Gln Asp Val     2921055  GCT GTG GAC TTC AGC CGG GAG GAG TGG GGG CTG CTG GGC CCG ACA CAG    1102293  Ala Val Asp Phe Ser Arg Glu Glu Trp Gly Leu Leu Gly Pro Thr Gln     3081103  AGG ACC GAG TAC CGC GAT GTG ATG CTG GAG ACC TTT GGG CAC CTG GTC    1150309  Arg Thr Glu Tyr Arg Asp Val Met Leu Glu Thr Phe Gly His Leu Val     3241151  TCT GTG GGG TGG GAG ACT ACA CTG GAA AAT AAA GAG TTA GCT CCA AAT    1198325  Ser Val Gly Trp Glu Thr Thr Leu Glu Asn Lys Glu Leu Ala Pro Asn     3401199  TCT GAC ATT CCT GAG GAA GAA CCA GCC CCC AGC CTG AAA GTA CAA GAA    1246341  Ser Asp Ile Pro Glu Glu Glu Pro Ala Pro Ser Leu Lys Val Gln Glu     3561247  TCC TCA AGG GAT TGT GCC TTG TCC TCT ACA TTA GAA GAT ACC TTG CAG    1294357  Ser Ser Arg Asp Cys Ala Leu Ser Ser Thr Leu Glu Asp Thr Leu Gln     3721295  GGT GGG GTC CAG GAA GTC CAA GAC ACA GTG TTG AAG CAG ATG GAG TCT    1342373  Gly Gly Val Gln Glu Val Gln Asp Thr Val Leu Lys Gln Met Glu Ser     3881343  GCT CAG GAA AAA GAC CTT CCT CAG AAG AAG CAC TTT GAC AAC CGT GAG    1390389  Ala Gln Glu Lys Asp Leu Pro Gln Lys Lys His Phe Asp Asn Arg Glu     4041391  TCC CAG GCA AAC AGT GGT GCT CTT GAC ACA AAC CAA GTT TCG CTC CAG    1438405  Ser Gln Ala Asn Ser Gly Ala Leu Asp Thr Asn Gln Val Ser Leu Gln     4201439  AAA ATT GAC AAC CCT GAG TCC CAG GCA AAC AGT GGC GCT CTT GAC ACA    1486421  Lys Ile Asp Asn Pro Glu Ser Gln Ala Asn Ser Gly Ala Leu Asp Thr     4361487  AAC CAA GTT TTG CTC CAC AAA ATT CCT CCT AGA AAA CGA TTG CGC AAA    1534437  Asn Gln Val Leu Leu His Lys Ile Pro Pro Arg Lys Arg Leu Arg Lys     4521535  CGT GAC TCA CAA GTT AAA AGT ATG AAA CAT AAT TCA CGT GTA AAA ATT    1582453  Arg Asp Ser Gln Val Lys Ser Met Lys His Asn Ser Arg Val Lys Ile     4681583  CAT CAG AAG AGC TGT GAA AGG CAA AAG GCC AAG GAA GGC AAT GGT TGT    1630469  His Gln Lys Ser Cys Glu Arg Gln Lys Ala Lys Glu Gly Asn Gly Cys     4841631  AGG AAA ACC TTC AGT CGG AGT ACT AAA CAG ATT ACG TTT ATA AGA ATT    1678485  Arg Lys Thr Phe Ser Arg Ser Thr Lys Gln Ile Thr Phe Ile Arg Ile     5001679  CAC AAG GGG AGC CAA GTT TGC CGA TGC AGT GAA TGT GGT AAA ATA TTC    1726501  His Lys Gly Ser Gln Val Cys Arg Cys Ser Glu Cys Gly Lys Ile Phe     5161727  CGG AAC CCA AGA TAC TTT TCT GTG CAT AAG AAA ATC CAT ACC GGA GAG    1774517  Arg Asn Pro Arg Tyr Phe Ser Val His Lys Lys Ile His Thr Gly Glu     5321775 AGG CCC TAT GTG TGT CAA GAC TGT GGG AAA GGA TTT GTT CAG AGC TCT    1822533  Arg Pro Tyr Val Cys Gln Asp Cys Gly Lys Gly Phe Val Gln Ser Ser     5481823  TCC CTC ACA CAG CAT CAG AGA GTT CAT TCT GGA GAG AGA CCA TTT GAA 1870549  Ser Leu Thr Gln His Gln Arg Val His Ser Gly Glu Arg Pro Phe Glu     5641871  TGT CAG GAG TGT GGG AGG ACC TTC AAT GAT CGC TCA GCC ATC TCC CAG    1918565  Cys Gln Glu Cys Gly Arg Thr Phe Asn Asp Arg Ser Ala Ile Ser Gln     5801919  CAC CTG AGG ACT CAC ACT GGC GCT AAG CCC TAC AAG TGT CAG GAC TGT    1966581  His Leu Arg Thr His Thr Gly Ala Lys Pro Tyr Lys Cys Gln Asp Cys     5961967  GGA AAA GCC TTC CGC CAG AGT TCC CAC CTC ATC AGA CAT CAG AGG ACT    2014597  Gly Lys Ala Phe Arg Gln Ser Ser His Leu Ile Arg His Gln Arg Thr     6122015  CAC ACC GGG GAG CGC CCA TAT GCA TGC AAC AAA TGT GGA AAG GCC TTC    2062613  His Thr Gly Glu Arg Pro Tyr Ala Cys Asn Lys Cys Gly Lys Ala Phe     6282063  ACC CAG AGC TCA CAC CTT ATT GGG CAC CAG AGA ACC CAC AAT AGG ACA    2110629  Thr Gln Ser Ser His Leu Ile Gly His Gln Arg Thr His Asn Arg Thr     6442111  AAG CGA AAG AAG AAA CAG CCT ACC TCA TAG CTC TCA AGC CAG TTG AAG    2158645  Lys Arg Lys Lys Lys Gln Pro Thr Ser ***                             6542159  AAA CCT TGC CTT TTC AGC TTG ACC CTG CAA TAT AAC ATG CAC AGG CCT    22062207  GCT TGT GAA TCA GGA CTG AAT GTG AAA GGG AAG TAT TGA GTG AGG ACA    22542255  TTC CCA AAA CCA AAG GAC AAC TGA GGA GAC TGC CCA GCA CAT AAT GAA    23022303  TAA ATA AGA AAA TGA GTG AGG AGT TAT TAA CAT CAT TTG GAA AAA AGA    23502351  TTT CCC ATT CAC TTG ATA TTG TTT GTT CAC TCA TTT AGT CAT TAA AAG    23982399  TGA GAT TAA TAA AAT CTG AAA ATG TTA TAT AAT AAC TTT AAA AAG CCA    24462447  GGT AAT TAA TAA TCT GCA CTG ATA TTA CAT CCA CAG TAC CAC AGT ATT    24942495  TAT GTG TAT GAA TTA AGG ATT AAA AGA TAA TGT GGA TAA AAA AAA AAA 25422543 AAA A 2546

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 (10)

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;

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.

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.

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.

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. 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.