CN1222935A - Compounds and method for immunotherapy and immunodiagnosis of prostate cancer - Google Patents

Abstract

Compounds and methods for treating and diagnosing prostate cancer are provided. The inventive compounds include polypeptides containing at least a portion of a prostate protein. Vaccines and pharmaceutical compositions for immunotherapy of prostate cancer comprising such polypeptides or DNA molecules encoding such polypeptides are also provided. The inventive polypeptides may also be used to generate antibodies useful for the diagnosis and monitoring of prostate cancer. Nucleic acid sequences for preparing probes, primers, and polypeptides are also provided.

Description

The compound and the method that are used for the immunotherapy and the immunodiagnosis of prostate cancer

Present invention relates in general to treatment of prostate cancer, diagnosis and supervision.More specifically the present invention relates to contain at least the polypeptide of the part of prostatein matter.Such polypeptide can be used for the treatment of the vaccine and the pharmaceutical composition of prostate cancer.This polypeptide also can be used for preparing as antibody etc. and be used for the diagnosis of the tumour that other of prostate cancer and patient may type and the compound that process monitors.

Prostate cancer is a modal cancer types among the male sex, estimates that in the male sex more than 50 years old sickness rate is 30%.A large amount of clinical evidence shows that human prostata cancers have the tendency of transferring in the bone, and this disease do not answer type like developing into male sex hormone from male sex hormone dependent form inevitably, thereby causes the increase of patient death rate.At present this ubiquity disease causes in the reason of cancer mortality second of row in the U.S. male sex.

Though have considerable research to be used for this treatment of diseases, still prostate cancer is difficult to treatment.Usually, treatment is based on operation and/or radiotherapy, but these methods are invalid in the case of suitable percentage ratio.Three kinds of prostate specific protein: prostate specific antigen (PSA) and prostatic acid phosphatase (PAP) have limited the ability of diagnosis and treatment.The level of PSA is not total consistent with existing of prostate cancer, and it can comprise in the benign prostatic hyperplasia (BPH) in the non-cases for prostate cancer of certain percentage ratio is positive.In addition, the observed value of PSA is relevant with prostatic volume but do not indicate the level of transfer.

Therefore, this area still needs to be used for the vaccine and the diagnostic method of the improvement of prostate cancer.

The invention provides the immunotherapy that is used for prostate cancer and the Compounds and methods for of diagnosis.On the one hand, the prostatein matter that includes the partial sequence of listing as SEQ ID No.2 and 4-8 or a kind of at least one immunogenicity polypeptide partly that this proteinic variant of the difference that conservative property replaces and/or modify is only arranged are provided.

In related fields, also provide polypeptide above the coding dna sequence dna, contain the expression vector of these dna sequence dnas and with these expression vector transfection or transformed host cells.In preferred embodiments, from the group of forming by intestinal bacteria (E.coli), yeast and mammalian cell, select host cell.

The present invention also provides and has included one or more SEQ ID No.1-8, and 20,21, the polypeptide of 25-31 or 44-57 or SEQ ID NO.9-19, the pharmaceutical composition of the nucleic acid of 22-24 or 32-43 and physiology acceptable carrier.The present invention also provides the vaccine that includes one or more such polypeptide or nucleic acid and nonspecific immune reaction toughener.

On the other hand, provide the method that suppresses prostate cancer development among the patient, comprised one or more SEQ ID No.1-8 that use effective dose to required patient, 20,21, the polypeptide of 25-31 or 44-57 or SEQ ID is No.9-19, the nucleic acid of 22-24 or 32-43.

Aspect another, the method that detects prostate cancer among the patient is provided, comprising: (a) available from patient's biological sample with can be incorporated into SEQ ID No 1-8,20,21, the binding reagents contact on the polypeptide of 25-31 or 44-57; (b) in the test sample with binding reagents bonded protein or polypeptide.

In related fields, the method that monitors prostate cancer development among the patient is provided, comprising: (a) available from patient's biological sample with can be incorporated into SEQ ID No 1-8,20,21, the binding reagents contact on the polypeptide of 25-31 or 44-57; (b) determine in the sample quantity with binding reagents bonded protein or polypeptide; (c) repeating step (a) and (b); And the amount of detected polypeptide in step (b) and (c) relatively.

In relevant range, the invention provides and aforementioned polypeptides bonded antibody preferred monoclonal antibody and the method that contains the diagnostic kit of such antibody and utilize such antibody inhibition prostate cancer to develop.

The present invention also provides the method that detects prostate cancer, comprising: (a) obtain biological sample from patient; (b) contact this sample with at least two Oligonucleolide primers in the polymerase chain reaction, have at least an Oligonucleolide primers to be specific to and be selected from SEQ ID No 9-19, the dna sequence dna of 22-24 and 32-43; (c) dna sequence dna that when having this Oligonucleolide primers to exist, increases in the test sample.In one embodiment, Oligonucleolide primers includes and is selected from SEQ ID No 9-19, a kind of dna sequence dna of 22-24 and 32-43 at least about 10 successive Nucleotide.

On the other hand, the invention provides the method for the prostate cancer that detects patient, comprising: (a) obtain biological sample from patient; (b) be selected from SEQ ID No.9-19 with being specific to, the oligonucleotide probe of the dna sequence dna of 22-24 and 32-43 contacts this sample; (c) in the test sample with the dna sequence dna of this oligonucleotide probe hybridization.In one embodiment, this oligonucleotide probe comprises and is selected from SEQ ID No.9-19, the dna sequence dna of 22-24 and 32-43 at least about 15 successive Nucleotide.

In case with reference to following detailed explanation and accompanying drawing, these or other aspect of the present invention is incited somebody to action apparent.All documents disclosed herein all with separately independently mode be incorporated by reference in this text and examine.

Fig. 1 shows the Western hybridization analysis available from the rat blood serum of using the immunity of rat prostate extract.

Fig. 2 shows the irreducibility SDS-PAGE of the rat immunity goods of Fig. 1.

Fig. 3 shows people's correspondence homologue of the protein-bonded supposition of a kind of rat kind sterol and combining of progesterone and estramustine.

As mentioned above, present invention relates in general to immunization therapy, diagnosis and supervision for prostate cancer Material and method. The material of invention is to contain to be at least a kind of demonstration and human prostate blood generally speaking The polypeptide that the part of immunoreactive human prostate protein is arranged clearly. The present invention also comprises and this The molecule of bright polypeptide combination (such as antibody or its fragment). Such molecule is called " in conjunction with examination herein Agent ".

Concrete, the invention discloses the human prostate that is provided by SEQ ID No.2 and 4-8 is provided Protein or only have conservative to replace and/or modify variant at least one of difference with this protein The polypeptide of part. " polypeptide " used herein word comprises any length amino of (containing full length protein) The acid chain, wherein amino acid residue links to each other by the covalency peptide bond. Therefore, include a kind of above-mentioned prostate The polypeptide of the part in the protein can be made up of this part fully, or this part be included in In the bigger polypeptide of appended sequence. Appended sequence can be derived from native protein or heterologous protein, And such sequence can be immunoreactivity and/or antigenic.

Human prostate protein used herein " immunogenicity part " is and is derived from trouble autoimmunity prostatitis The people's of adenositis seroreaction or can with the serum that is derived from the prostatitic rat model of autoimmunity The part of reaction. In other words, immunogenicity part can cause a kind of immune response and can be incorporated into On the antibody that exists in the prostatitis serum. The autoimmunity prostatitis can, for example, by annotating Penetrating BCG vaccine (BCG) (a kind of avirulent Mycobacterium bovis (Mycobacterium bovis)) controls Take place after treating carcinoma of urinary bladder. In the prostatitic rat model of autoimmunity, the usefulness rat prostate Detergent extract immune rat. The serum in arbitrary source can be used for described herein in these sources Derive polypeptide reaction of human prostate. Antibody can utilize such as those of ordinary skills in conjunction with measuring Known either method is carried out, for example, at Harlow and Lane, " antibody: laboratory hand Volume " (Antibody:A Laboratory Manual) cold spring harbor laboratory, the cold spring port, New York, Described in 1988. For example, polypeptide can be fixed on solid support (as following) also and the patients serum connects Touch so that intraserous antibody is combined with immobilized polypeptide. Remove unconjugated serum, with as¹²⁵I The albumin A of mark detects the antibody of combination.

" variant " described herein only has conservative to replace and/or modify difference with described polypeptide Polypeptide, thereby immunotherapeutical, the antigenicity of this polypeptide or the molecule of being combined with this polypeptide and/or examine Disconnected character all keeps. For the prostatein matter that immune response character is arranged, usually above-mentioned by modifying The immunoreactivity of the polypeptide of estimating in the lump this modified of peptide sequence and identify variant. Produce being used for Give birth to the prostatein matter of diagnostic binding reagents, can pass through the generation inspection of the polypeptide of evaluation modified Survey has or not the ability of the antibody of prostate cancer to identify a kind of variant. For example, can be with described herein Exemplary process preparation and the sequence of testing this modified.

" conservative replacement " used herein word refers to that a seed amino acid is by the another kind that close character is arranged 49-Phe ,82-Ser,115-Arg,144-Met,145-Asn ,161-Arg,169-Met Human Connective tissue growth factor, thus chemistry of peptides field those of ordinary skill be contemplated to this polypeptide secondary structure and Hydrophilic nmature does not change basically. Usually, following amino acid group represents conservative and changes: (1) ala, Pro, gly, glu, asp, gln, asn, ser, thr; (2) cys, ser, tyr, thr; 3) val, ile, leu, Met, ala, phe; (4) lys, arg, his and (5) phe, tyr, trp, his.

In addition, variant also can contain other to be modified, and comprises antigenic property, secondary knot to this polypeptide Structure and hydrophilic nmature have amino acid whose deletion or the interpolation of minimum influence. For example, a polypeptide can be even Link on signal (or guide) sequence of N end of this protein, it instructs simultaneously or after the translation in translation The transfer of this protein. This polypeptide also can be coupled to a joint or be convenient to the synthetic, pure of this polypeptide Change or other sequence of identification on (such as polyhistidyl), or be attached to and consolidate to increasing this polypeptide by coupled On the sequence of phase holder. For example, a polypeptide can be by coupled to an immunoglobulin fc region territory On.

Can be from suitable PC-3 such as Ln Cap.fgc (ATCC No.1740-CRL) separate in and have the 1-8 by SEQ ID No, 20,21 and one of the sequence that provides of 25-31 Polypeptide. Ln Cap.fgc is a kind of representative prostate gland cancer cell of particularly preferred human prostata cancer System. As people's cancer, the heterograft in the SCID mouse of Ln Cap.fgc cell forms continuous life Long tumour is replied testosterone, secretion PSA and reply the existence of marrow composition (such as transferrins). Tool Body ground, this polypeptide can with as the people such as Sambrook " molecular cloning " (Molecular Cloning: A Laboratory Manual) cold spring harbor laboratory, the cold spring port, New York is (with the literary composition of wherein quoting And method described in detail below, by expressing employment prostatitis serum screening Ln Cap. The fgc cDNA library separates. SEQ ID No.48 and 49 polypeptide can be by discussing with top The prostatitic rat model of autoimmunity in serum screening separate from Ln Cap/fgc clone Arrive. The polypeptide of SEQ ID No.50-56 can be with passing through the people prostatitis such as the method that describes in detail among the embodiment 4 The adenositis serum screening separates from Ln Cap/fgc clone. The polypeptide of SEQ ID No.44-47 can Separate from people's seminal fluid with the method that describes in detail such as embodiment 2. In case obtain the DNA order of coded polypeptide Row, any above-mentioned modification all can utilize the standard sudden change such as the mutation site-specific of oligonucleotides guiding Technology and introducing easily.

Polypeptide disclosed herein also can produce by synthetic or recombination method. Less than about 100 ammonia What base was sour usually can use those of ordinary skills less than the synthetic of 50 amino acid whose polypeptide Known technology produces. For example, such polypeptide is synthetic can be with arbitrary commercial possible solid phase Technology as: the Merrifield solid-phase synthesis is finished, and wherein amino acid is joined growth in proper order On the amino acid chain. See Merrifield Journal of the American Chemical Society (J.Am.Chem. Soc.) 85:2149-2146,1963. The automatic pressing forming apparatus of polypeptide can be from such as Applied The supplier of BioSysterms company (Foster City, CA) obtains, and by manufacturer's indication operation.

In addition, any aforementioned polypeptides all can be inserted into table by the dna sequence dna of this polypeptide of will encoding Reach carrier and in suitable host, express the method recombinant production of this albumen. Any this area is common The expression vector that the technical staff knows all can be used for expressing recombinant polypeptide of the present invention. Expression can be in office What suitable usefulness contains the expression vector conversion of dna molecular of the recombinant polypeptide of encode or the host of transfection Realize in the cell. Suitable host cell comprises prokaryotic, yeast and higher eucaryotic cells. Excellent The host cell that choosing utilizes is for Escherichia coli, yeast or such as the mammalian cell of Chinese hamster ovary celI System. The dna sequence dna of expressing the by this way naturally occurring polypeptide, naturally occurring of can encoding The part of polypeptide or its other variant.

Usually, no matter its preparation method how, polypeptide disclosed herein is with basically pure form system (that is: form with amino acid and this polypeptide of Primary Structure Analysis is homogeneous). Preferably, this polypeptide Purity is at least 90%, and preferred purity is at least 95%, and most preferred is purity at least 99%. In specific preferred embodiment (being described in further detail below), pure polypeptide is added in basically Be used for one or more methods disclosed herein in pharmaceutical composition or the vaccine.

The polypeptide of the present invention that contains an immunogenicity part of prostatein can be used for the immunotherapy of prostate cancer, and wherein this polypeptide stimulates patient's self the immunne response at prostate tumor cells.On the other hand, the invention provides and utilize SEQ ID No.1-8,20,21, one or more immunoreactivity polypeptide of 25-31 and 44-57 (or DNA of such polypeptide of encoding) are used for the method for the immunotherapy of patient's prostate cancer." patient " used herein is meant any warm-blooded animal, preferred people.Patient can be subjected to a kind of puzzlement of disease, and also can not having can detected disease.Therefore, above-mentioned immunoreactivity polypeptide can be used for treating the development of prostate cancer or inhibition prostate cancer.Can operation remove primary tumo(u)r and/or with the treatment of radiotherapy and traditional chemotherapeutics before or after use this polypeptide.

In these areas, this polypeptide is generally comprised among pharmaceutical composition and/or the vaccine.Pharmaceutical composition can comprise one or more polypeptide, and every kind all contains one or more above-mentioned sequences (or its variant) and physiology acceptable carrier.Vaccine can comprise one or more such polypeptide and nonspecific immune reaction toughener as a kind of adjuvant, biodegradable microsphere (as: poly(lactic acid) glycolide) or liposome (this polypeptide is integrated in wherein).Pharmaceutical composition or vaccine also can contain antigenic other epi-positions of prostatic cell, these epi-positions or be integrated into the combination polypeptide single polypeptide of a plurality of epi-positions (that is: contain) or be the isolated polypeptide form.

In addition, pharmaceutical composition or vaccine can contain the DNA of one or more aforementioned polypeptides of encoding, thereby this polypeptide produces on the spot.In such pharmaceutical composition and vaccine, this DNA can exist in the drug delivery system that any those skilled in the art know, and comprises expression of nucleic acid system, bacterium and expressing viral system.Suitable expression of nucleic acid system is included in and expresses necessary dna sequence dna (as suitable promotor) among the patient.The bacterium drug delivery system comprises a kind of bacterium (as bacill calmette-guerin) in the antigenic epi-position of its cell surface expression prostatic cell of use.In preferred embodiments, can utilize expressing viral system (as: cowpox or other poxvirus, retrovirus or adenovirus), comprising utilizing non-virulent (defective type), having the virus of replication to import this DNA.Suitable system is disclosed, as people such as Fisher-Hoch, PNAS 86:317-321,1989; People such as Flexner, Ann.N.Y.Acad.Sci.569:86-103,1989; People such as Flexner, vaccine (Vaccine) 8:17-21,1990; U.S. Patent No. 4,603,112,4,769,330 and 5,017,487; WO 89/01973; U.S. Patent No. 4,777,127; GB2,200,651; EP0,345,242; WO 91/02805; Berkner, biotechnology (Biotechniques) 6:616-627,1988; People such as Rosenfeld, science 252:431-434,1991; People such as Kolls, PNAS 91:215-219,1994; People such as Kass-Eisler, PNAS 90:11498-11502,1993; People such as Guzman, circulation (Circulation) 88:2838-2848,1993 and people such as Guzman, Cir.Res.73:1202-1207,1993.The scheme that DNA is incorporated in these expression systems is known for those of ordinary skills.This DNA also can be " exposing ", as people such as disclosed PCT application WO 90/11092 and Ulmer, the described and Cohen science 259:1691-1692 of science 259:1745-1749,1993 summaries.The DNA bag can be increased the absorption of naked DNA to the biodegradable pearl that can effectively be transferred to cell.

The approach of medication, number of times and dosage can be with individual different the variations, also can be with reference at present in the method for exempting from use in the disease treatment of other disease.Usually, pharmaceutical composition and vaccine can be by injection (as: intracutaneous, intramuscular, intravenously or subcutaneous), nasal cavity (as: by air-breathing) or oral administrations.The sustainable 3-24 time-of-week of being longer than of the 1st to the 10th dose administration.Preferred use 4 doses, medication can regularly be strengthened thereafter in every dose of interval 3 months.Also can use other scheme for single patient.Proper dosage is can effectively improve in subject patient at the polypeptide of the immunne response of prostate tumor cells or the amount of DNA.Suitable immunne response is wanted high 10-50% than baseline values (as: untreated) at least.Usually, in the potion (or by DNA produce in situ in the potion) scope of the amount of polypeptide from about 1pg to about 100mg per kilogram host, typically from about 10pg to about 1mg, preferably from about 100pg to about 1 μ g.The volume of proper dosage changes with patient's size, and typically scope is from about 0.01 milliliter to about 5 milliliters.

Though the suitable carriers that any those of ordinary skills know all can be used in the pharmaceutical composition of the present invention, the type of carrier is then different according to the difference of administering mode.For parenterai administration, as subcutaneous injection, preferred carrier comprises water, physiological saline, ethanol, fat, wax and/or damping fluid.For oral administration, any above-mentioned carrier or solid phase carrier such as: N.F,USP MANNITOL, lactose, starch, Magnesium Stearate, soluble saccharin, talcum, Mierocrystalline cellulose, glucose, sucrose and/or magnesiumcarbonate all can use.Biodegradable microsphere (as the poly(lactic acid) glycolide) also can be as the carrier of pharmaceutical composition of the present invention.Suitable biodegradable microsphere has been disclosed in as U.S. Patent No. 4,897, in 268 and 5,075,109.

Many nonspecific immune reaction tougheners all can be used for vaccine of the present invention.For example can comprise a kind of adjuvant.Comprise the material that is used to prevent that antigen from decomposing rapidly in most of adjuvants, as aluminium hydroxide or mineral oil and as the nonspecific immune reaction stimulator of liposome A, Bordetella pertussis (Bordella pertussis) or mycobacterium tuberculosis (Mycobacterium tuberculosis).Such adjuvant can buy, as: freund 's incomplete adjuvant and Freund's complete adjuvant (Difco Laboratories, Detroit, MI) and the Merck adjuvant 65 (Merck and Company company, Rathway, NJ).

Polypeptide disclosed herein also can be used for the stripped treatment of prostate cancer.For example, can utilize business-like cellular segregation system as: CellPro Incorporated ＇ s (Bothell, WA) U.S. Patent No. 5,240 (is seen, 856 by the CEPRATETM system; 5,215,926; WO89/06280; 91/16116 and 92/07243) from patient's peripheral blood, separates immune system cell such as the T cell.Isolated cells stimulates so that T cells with antigenic specificity to be provided with one or more immuno active polypeptides that are included in as in the microsphere.Utilize standard technique to make specific for tumour antigen T cell mass amplification and these cells are got back among the patient.

In addition, polypeptide of the present invention also can be used to produce such as antibody or its segmental binding reagents that can detect the metastatic human tumor of prostate.

Binding reagents of the present invention can make by the known method of the those of ordinary skills that comprise exemplary process described herein.Binding reagents can utilize representative authentication method described herein to distinguish the patient who has and do not have prostate cancer.In other words, appearance at the antibody of prostatein or its suitable part or other binding reagents will produce the signal that the patient Central Plains at least about 20% is sent out or metastatic prostate cancer exists that an indication is subjected to this disease puzzlement, and produce an indication do not have former or metastatic prostate cancer at least about 90% individuality in do not have the signal that this disease exists.The suitable fragments of this prostatein is meant such fragment, it can produce a kind of all (at least about 80% basically, preferably at least about 90%) binding reagents that utilizes former of indication among the patient of full-length proteins mass-energy indication prostate cancer or metastatic prostate cancer to exist, indicate in can also be when the useful full length protein test of the institute basically negative sample not have prostate cancer.The representativeness of narrating below identifies that (as: double antibodies sandwich evaluation) can be used for estimating the ability that binding reagents detects the metastatic human tumor of prostate.

As described here the polypeptide of preparation produce the ability detect former or metastatic human tumor of prostate generally can be by generation at one or more antibody (as utilizing representational method described herein) of this polypeptide and measure this antibody detects this tumour in patient ability and estimate.This mensuration can be tested and appraised to be derived from and not have in patient's the biological sample of former or metastatic prostate cancer and finish with the existence of the polypeptide of the antibodies that is produced.Such evaluation can be finished as utilizing the exemplary process of narrating below.The polypeptide that produces the antibody can detect at least 20% former or metastatic prostate tumour in this way is considered to produce the antibody that can detect former or metastatic human tumor of prostate.Polypeptid specificity antibody can separately or be united use to increase susceptibility.

The polypeptide of former of energy detection or metastatic human tumor of prostate can be as the mark of the progression of disease of diagnosing patient's prostate cancer or supervision patient.In one embodiment, patient's prostate cancer can be diagnosed with respect to the level of predetermined threshold value available from one or more aforementioned polypeptides in patient's the biological sample by estimating.As used herein, suitable " biological sample " comprises blood, serum, urine and/or prostatic secretion.

Can utilize any binding reagents that is specific to this (a bit) polypeptide to estimate the level of one or more aforementioned polypeptides.A kind of in the present invention " binding reagents " is and the above-mentioned any reagent of polypeptide bonded (as a kind of compound or cell)." combination " used herein is meant the non-covalent combination between two isolating molecules (each all be free (that is: in solution) or on cell or solid support surface), so form " mixture ".Such mixture can be the surface that free or (covalently or non-covalent ground) are fixed on support material.The bonded ability is generally estimated by the binding constant of measuring the mixture that forms.Binding constant is the numerical value of complex concentration divided by the product acquisition of concentration of component.Usually, work as the binding constant of the mixture that forms in the present invention above 10 ³These two kinds of compounds are called " combination " during L/mol.Can utilize the known method of those of ordinary skills to measure binding constant.

Any reagent that satisfies above-mentioned requirements all can be binding reagents.For example, binding reagents can be the rrna that has or do not have peptide composition, a kind of RNA molecule or peptide.In an embodiment preferred, binding partner is a kind of antibody or its fragment.This (a bit) antibody can be polyclone or monoclonal.In addition, antibody can be strand, chimeric, the CDR transplanting or humanized.Can with narration herein or method that other those of ordinary skills know prepare antibody.

The authentication method that has many those of ordinary skills to know is used for the polypeptide marker with the binding partner test sample.See as Harlow and Lane antibody: laboratory manual (Antibodies:A Laboratory Manual) cold spring harbor laboratory, 1988.In an embodiment preferred, this evaluation comprises uses the binding partner that is fixed on the solid support to go to separate in conjunction with polypeptide and with the rest part of sample.The bonded polypeptide can detect with the secondary binding partner that contains reporter group again.Suitable secondary binding partner comprises and binding partner/polypeptide complex bonded antibody.In addition, can utilize competitive the evaluation, wherein it is attached on the immobilized binding partner with a reporter gene labeling polypeptide and after binding partner and sample are hatched.Sample composition suppresses labeling polypeptide and binding partner bonded degree is reactive indication of this sample and immobilization binding partner.

Solid support can be any material that antigen that those of ordinary skills know can adhere to.For example: solid support can be sample well or nitrocellulose membrane or other suitable membrane in the microwell plate.In addition, upholder can be an a pearl or garden dish, as glass, glass fibre, latex or as the plastic material of polystyrene or polyvinyl chloride.This upholder also can be magnetic-particle or Fibre Optical Sensor, and as U.S. Patent No. 5,359,681 is disclosed.The several different methods that is described in detail in patent and scientific literature that can utilize those skilled in the art to know is fixed to binding reagents on the solid support.In the present invention, " fix " speech and refer to non-covalent combination (as absorption) and covalently bound (can be antigen with functional group on the upholder directly be connected or can be connection) by the linking agent mode.Preferably by fixing on the hole that is adsorbed onto microwell plate or on the film.In this case, absorption can be finished by contact one suitable period of binding reagents on solid support in suitable damping fluid.Change along with temperature variation duration of contact, but typically between about 1 hour to about 1 day.Usually, the binding reagents of q.s is fixed with regard to being enough to the hole (as polystyrene or polyvinyl chloride) of scope from about 10ng to the preferably about 100ng of about 10 μ g to the binding reagents contact plastic microporous plate of about 1 μ g amount.

Usually, by at first with can with upholder and as hydroxyl or amino binding reagents on the difunctionality reagent that reacts of functional group and upholder react and realize binding reagents covalently bound on solid support.For example, binding reagents can utilize benzoquinones covalently to be connected on the upholder of suitable polymer coating, also can covalently boundly (see by finishing with the aldehyde radical condensation on the amino on the binding partner and active hydrogen and the upholder, as Pierce immunological technique catalogue and handbook, 1991, A12-A13)

In specific embodiments, evaluation is that double antibodies sandwich is identified.This authentication method at first contacts the antibody that has been fixed on the solid support (being generally the hole of microwell plate) with sample, and the polypeptide in the sample is attached on the immobilized antibody.Unconjugated sample separates with immobilized polypeptide-antibody complex, and adding can be in conjunction with two anti-(the containing reporter group) in the other site of this polypeptide again.The method that utilization is suitable for the specificity reporter group detects the anti-amount of two on the solid support that is retained in.

More specifically, in case as above-mentioned antibody is fixed on the upholder, remaining protein binding site will be closed on the upholder.Suitable encapsulant such as bovine serum albumin or the Tween 20 that any those of ordinary skills know ^TM(Sigma chemical company, St.Louis MO) all can use.Immobilized antibody and sample are hatched together, and polypeptide is attached on the antibody.Can be with suitable diluent such as phosphate-buffered saline (PBS) dilute sample before hatching.Usually, be the time that is enough to detect available from the existence of this polypeptide in prostate cancer patient's the sample suitable duration of contact (that is: incubation time).Preferably, duration of contact be enough to realize reaching at least equilibrium level between combination and the unconjugated polypeptide 95% in conjunction with level.Those of ordinary skills know that reaching the required time of balance can be tested and appraised through easily detecting in conjunction with level of taking place after a while.Under the room temperature, general about 30 minutes incubation time is just enough.

Can by with suitable as containing 0.1%Tween20 ^TMThe damping fluid washing solid support of PBS remove unconjugated sample.Two anti-the joining on the solid support that to contain reporter group then.Preferred reporter group comprises enzyme (as horseradish peroxidase), substrate, cofactor, inhibitor, dyestuff, radionuclide, luminophore, fluorophor and vitamin H.Can realize the coupling of antibody and reporter group with the standard method that those of ordinary skills know.

Two anti-hatch for some time that is enough to detect the bonded polypeptide with immobilized antibody-polypeptide complex.Can determine the suitable time in conjunction with level by what take place after detection for some time.Removing unconjugated two resists and resists with reporter group detection bonded two.The method that is used for the examining report group depends on the character of reporter group.Generally be suitable for scintillation counting or radioautograph method for the radioactivity group.Can utilize optical means to detect dyestuff, luminophore and fluorophor.Can be with the avidin detection of biological element that is coupled to different reporter group (being generally radioactivity or fluorophor or enzyme).Generally detect the enzyme reporter group with optics or other methods analyst reaction product subsequently by adding substrate (generally through after a while).

For the existence that detects prostate cancer whether, detect from the signal that is retained in the reporter group on the solid support and generally can compare with the corresponding signal of predetermined threshold value.In an embodiment preferred, threshold value is the mean value of the signal that records when hatching from the patient's of no prostatitis gland cancer sample when immobilized antibody.Usually, the sample that produces the signal that is higher than three standard deviations of predetermined threshold value is thought the prostate cancer positive.In another embodiment preferred, utilize to receive and handle the method (clinical epidemiology: the basic science of clinical medicine (Clinical Epidemiology:A Basic Science for ClinicalMedicine) Little Brown and Co. of curve (Receiver Operator Curve) according to people such as Sackett, 1985,106-7 page or leaf) determines threshold value.In brief, in this embodiment, can be used for the True Positive Rate (being susceptibility) of the possible threshold value of diagnostic test results and the curve of false positive rate (100%-specificity) is measured corresponding to each from paired.Be threshold value the most accurately near the threshold value in the curve of the upper left corner value of maximum area (that is: comprise), the sample that produces the signal that is higher than the threshold value that this method records can be thought the positive.In addition, threshold value can be moved to the left along curve, to reduce false positive rate or to move right, to reduce false negative rate.Usually, the sample that produces the signal be higher than the threshold value that this method records can be thought the prostate cancer positive.

In relevant embodiment, can be with flowing or the band method of testing, wherein antibody is fixed on as on the nitrocellulose filter.In flowing test, when sample passed film, the polypeptide in the sample was attached on the immobilized antibody.Cross two anti-just being attached on the antibody-polypeptide mixture that the film tense marker is crossed when containing two anti-solution stream.It is anti-to detect bonded two with aforesaid method.In the band method of testing, an end that is combined with the film of antibody is immersed in the solution that contains sample.Sample passes along film and contains the zone that two anti-zones move to immobilized antibody.Indicate the concentration of prostate cancer in two of the immobilized antibody zone anti-concentration.Typically, two anti-concentration produce a kind of pattern of as the linearity that can with the naked eye see in such site.No such pattern indication negative findings.Usually, the amount of selecting to be fixed on the antibody on the film make it can generation can with the naked eye pick out in contain the method for discussing useful front and be enough to produce the biological sample of polypeptide level of positive signal in the double antibodies sandwich evaluation pattern.Preferably, the scope of amount that is fixed on the antibody on the film from about 25ng to about 1 μ g, preferred from about 50ng to about 500ng.Typically can finish such test with very small amount of biological sample.

Certainly, antigen of the present invention and antibody are applicable in existing various other authentication method.Top description only is intended to for example.

In another embodiment, top polypeptide can be as the mark of prostate cancer development.In this embodiment, after the above-mentioned authentication method that is used for the diagnosis of prostate cancer carries out for some time, the variation of the level of evaluation response polypeptide.For example, can measure once in every 24-72 hour, continue 6 months to 1 year, carry out where necessary thereafter.Usually, develop in those prostate cancers in the each patient who all increases of the polypeptide level that binding reagents detects.On the contrary, prostate cancer does not develop when the level of reactive polypeptide maintains an equal level or descends in time.

The antibody that is used for top method can make with any of the known the whole bag of tricks of those of ordinary skills.See as Harlow and Lane, " antibody: laboratory manual ", cold spring harbor laboratory, 1988.In a kind of such technology, the immunogen that will include antigenic polypeptide initially is expelled to multiple mammiferous in any (as: mouse, rat, rabbit, sheep and goat).In this step, polypeptide of the present invention can be not modified as immunogen.In addition, the polypeptide for short relatively can cause stronger immune response if this polypeptide and carrier proteins (as bovine serum haemproteins or keyhole limpet hemocyanin) are united use.Immunogen is expelled among the animal host, preferably according to the predetermined scheme of one or many booster immunization associating, regularly animal is got blood.By as: utilize method purifying from antiserum(antisera) of the affinity chromatography be coupled to the polypeptide on the suitable solid phase upholder to be specific to the polyclonal antibody of this polypeptide.

Can be for example: utilize Kohler and Milstein, European Journal of Immunology (Fur.J.Immunol.) 6:511-519,1976 the method and the preparation of improving one's methods thereof are specific to the monoclonal antibody of target antigen polypeptide.In brief, these methods comprise that preparation can produce the immortal cell line of the specific antibody with expectation.This clone can be from for example available from producing through the splenocyte of animal of immunity as mentioned above.Splenocyte by as with preferably follow immune animal homologous myeloma cell fusion partner merge and immortalization.Can utilize multiple integration technology.For example: can support the hybrid cell growth but do not support on the selective medium that the myeloma cell grows with being inoculated into lower concentration after splenocyte and myeloma cell and a kind of non-ionic detergent mixed for several minutes.Preferred selection technology is to utilize HAT (xanthoglobulin, aminopterin, thymidine) to select.Through behind the time enough, be generally about 1 to 2 week, can be observed the clone of hybridization.The combination at this polypeptide of selecting and test single clone is active.Preferably have hyperergy and specific hybridoma.

Can from growth hybridoma clone's supernatant liquor, separate monoclonal antibody.In addition, can utilize different technologies to increase output, as: hybridoma cell line is expelled in the abdominal cavity of suitable vertebrates such as mouse.Can from ascites or blood, gather in the crops monoclonal antibody then.Can by routine techniques as: the pollutent in the antibody is removed in chromatography, gel-filtration, precipitation and extraction.Polypeptide of the present invention also can be used for the purification process as the affinity chromatography step.

Monoclonal antibody of the present invention also can be as therapeutical agent to dwindle or to eliminate tumor of prostate.Antibody can use (for example suppressing to shift) also can use together with one or more therapeutical agents separately.Suitable therapeutical agent described herein comprises radionuclide, differentiating inducer, medicine, toxin and derivative thereof.Preferred nucleic comprises ⁹⁰Y ¹²³I, ¹²⁵I, ¹³¹I, ¹⁸⁶Re, ¹⁸⁸Re, ²¹¹At and ²¹²Bi.Preferred medicine comprises methotrexate and pyrimidine and purine analogue, and preferred differentiating inducer comprises Buddhist ripple ester and butyric acid.Preferred toxin comprises ricin, toxalbumin, diphtheria toxin (diptheria toxin), Toxins,exo-, cholera, gelonin, false unit cell extracellular toxin, shiga toxin and Pokeweed antiviral protein.

Therapeutical agent can with directly or indirectly (as the passing through linking group) coupling (as covalently bound) of suitable monoclonal antibody.When all have a kind of substituting group can and just may be between antigen and antibody during other substitution reaction direct reaction.For example: a kind of nucleophilic group such as amino or sulfydryl, on the one hand can with the group that contains carbonyl such as a kind of acid anhydrides or carboxylic acid halides reaction; Can react with the alkyl that contains good leavings group (as halogenide) on the other hand.

In addition, can expect between therapeutical agent and the antibody by the linking group coupling.Thereby linking group can be used as spacer and antibody and reagent is kept at a certain distance away avoid interference to binding ability.Linking group also can be used for increasing the chemical reactivity of substituting group on reagent or antibody, thereby increases link coupled efficient.Chemically reactive increase also can promote to use impossible originally reagent that utilizes or the functional groups on the reagent.

Conspicuous to those skilled in the art have different difunctional or poly functional reagents, comprises (as Pierce chemical company, Rockford is described in the catalogue of IL) of of the same race or xenogenesis function, can be used as linking group.For example can carry out coupling by the saccharide residue of amino, carboxyl, sulfydryl or oxidation.There is a large amount of reference to describe this method, as people's such as Rodwell U.S. Patent No. 4,671,958.

For more efficiently therapeutical agent when not having immune link coupled antibody moiety of the present invention, can expect to use a kind of linking group that in entering cell processes or after entering cell, can cut.Existing a series of different linking groups that cut are described.The mechanism that reagent discharges in these linking groups born of the same parents comprises that reduction by disulfide linkage is (as the U.S. Patent No. 4 of Spitler, 489,710), the irradiation of photo-labile key (as: people's such as Senter U.S. Patent No. 4,625,014), the hydrolysis of derivative amino side chain (as: people's such as Kohn U.S. Patent No. 4,638,045), the hydrolysis of blood plasma complement-mediated (as: people's such as Rodwell U.S. Patent No. 4,671,958) and the method for acid catalyzed hydrolysis (as: people's such as Blattler U.S. Patent No.s 4,569,789) cutting.

Sometimes expect plurality of reagents is coupled on the antibody.In one embodiment, a kind of a plurality of molecules of reagent are coupled on the antibody molecule.In another embodiment, polytype reagent can be coupled on a kind of antibody.No matter specific embodiment is how, can be by the immune conjugate of different methods preparation with plurality of reagents.For example: plurality of reagents can direct and a kind of antibody molecule coupling, perhaps can utilize the joint that a plurality of attachment sites can be provided.In addition, also can utilize carrier.

Carrier can multiple formation comprise reagent, comprises directly or the covalent bonds by linking group.Suitable carriers comprises protein such as albumin (as: people's such as Kato U.S. Patent No.s 4,507,234), peptide and polysaccharide such as glycosaminoglycan (as: people's such as Shih U.S. Patent No.s 4,699,784).Carrier also can comprise reagent as methods such as (as U.S. Patent No.s 4,429,008 and 4,873,088) in liposome vesicle by non covalent bond or by encapsulated.The carrier that is specific to radionuclide comprises: the small molecules of radioactive halogenation and huge legendary turtle combination compound.For example: U.S. Patent No. 4,735,792 disclose the small molecules of representative radioactive halogenation and synthesizing of theirs.The huge legendary turtle compound of radionuclide can be from comprising that containing nitrogen and sulphur atom closes mixture as the huge legendary turtle of the donor atom that is used for bond or metal oxide, nucleic and form.For example: people's such as Davison U.S. Patent No. 4,673,562 disclosed representative huge legendary turtle combination compounds and theirs synthetic.

Can use different routes to be used for the administration of antibody and immune conjugate.Typically, administration is adopted intravenously, intramuscular, subcutaneous mode or in the implantation position administration of the tumour of excision.Obviously, the exact dosage desired of antibody/immune conjugate depends on the clearance rate of antigenic density and antibody on employed antibody, the tumour.

Diagnostic reagent of the present invention also can comprise the dna sequence dna of one or more aforementioned polypeptides of coding or its one or more parts.For example: can utilize two Oligonucleolide primers to be derived from the tumor of prostate specificity cDNA of biological sample with amplification at least in based on the evaluation of polymerase chain reaction (PCR), wherein at least one this Oligonucleolide primers is specific to the dna molecular of code book invention polypeptide.Utilize the existence of the cDNA of known technology in this area such as detected through gel electrophoresis amplification.Similarly, the oligonucleotide probe that in hybridization is identified, can utilize the dna molecular that is specific to code book invention polypeptide with the detection of biological sample in the existing of invention polypeptide.

" being specific to the Oligonucleolide primers/probe of dna molecular " used herein is meant and have an appointment at least oligonucleotide sequence of 80% homology of target DNA molecule, preferably at least about 90%, more preferably at least about 95% homology.The Oligonucleolide primers and/or the probe that can be used for diagnostic method of the present invention preferably contain at least about 10-40 Nucleotide.In an embodiment preferred, Oligonucleolide primers comprises the dna molecular of a coding peptide species disclosed herein at least about 10 successive Nucleotide.Preferably, the oligonucleotide probe that is used for diagnostic method of the present invention comprise a coding peptide species disclosed herein dna molecular at least about 15 successive Nucleotide.Be used for the evaluation of PCR-based and technology that hybridization is identified and be that this area is known (sees that the source is the same as people such as Mullis; Ehrlich, the source is the same).Primer and probe thereby can be used for being preferably blood, seminal fluid or prostate gland and/or prostate tumor tissue and detect prostate gland and/or tumor of prostate sequence at biological sample.

Provide the following example unrestricted in order to illustrate.Embodiment 1A. utilizes the scorching serum of human prostate isolated polypeptide from Ln Cap.fgc

By utilizing the scorching serum of human prostate to separate representative polypeptide of the present invention with following method screening PC-3.By synthetic from the mRNA reverse transcription of PC-3 Ln Cap.fgc (ATCCNo.1740-CRL) from purifying, cDNA clone with gained is inserted into λ ZAP II (Stratagene again, La Jolla, method CA) makes up human prostata cancer cDNA expression library.

The scorching serum of human prostate is diagnosed as available from one and takes the prostatitic patient of BCG treatment bladder cancer future trouble autoimmunization.With as people such as Sambrook, " molecular cloning " (Molecular Cloning:A laboratory Manual), cold spring harbor laboratory, the cold spring port, NY, the method described in 1989 is with this serum screening Ln Cap cDNA library.Particularly, with about 10 ⁴The Ln CapcDNA library coating LB flat board of pfu is cultivated the die of plaque first that obtains after 4 hours on the nitrocellulose filter that isopropylthio-beta galactose glycosides (IPTG) soaks into down for 42 ℃.Dull and stereotyped 42 ℃ were continued cultivation 5 hours down and obtain plaque die for the second time by 37 ℃ of following incubated overnight.Filter membrane (contains 1%Tween20 with PBS-T washing 3 times with PBS ^TM) sealed 1 hour, once more with PBS-T washing 3 times, then under agitation with the scorching serum of the human prostate that diluted with 1: 200 overnight incubation together.Wash filter membrane 3 times with PBS-T then, and under agitation cultivated 1 hour with the albumin A (1 μ l/15ml PBS-T) of 125I mark.Filter membrane to exposure from 16 hours to 7 days the different time in the scope.The plaque that signal is arranged on the duplicate impression by renewed vaccination to the LB flat board.The gained plaque is handled filter membrane with the same method on the filter membrane that duplicates.These filter membranes 4 ℃ times and the scorching serum of human prostate (dilution in 1: 200) stir culture are spent the night.By with filter membrane to exposure in 16 hours to 11 days the scope different time with as above-mentioned ¹²⁵The albumin A of I mark is observed positive plaque.Finish according to manufacturers indication in positive human prostatitis antigen cDNA clone's the body and cut out.B. the sign of polypeptide

(Foster City utilizes on CA) forward and reverse primer to obtain the dna sequence dna of positive colony in the automatic sequencer 373A of Applied Biosystems company type.After this cDNA sequence of coding institute isolated polypeptide is called HPA8, HPA13, HPA15～HPA17, HPA20, HPA25, HPA28, HPA29, HPA32～HPA38 and HPA41 are listed in SEQ ID No.32 and 33 respectively, 34 and 35,36,9 and 10,11,12,13 and 14,15,37 and 38,16,39,22 and 23,17 and 18, in 19,24,40 and 41,42 and 43.The 3 ＇ sequences of HPA16 and HPA20 are identical.HPA13, HPA16, HPA20, HPA29 and HPA33 are considered to have the eclipsed clone of 5 new ＇ end points.Two positive colonies are identified with HPA15 identical.Equally, HPA15, HPA34 and HPA37 also find it is overlapping clone.Based on the cDNA sequence of in framework, determining with the N-end parts of beta-galactosidase enzymes (LacZ), isolated polypeptide HPA16, HPA17, HPA20, HPA25, HPA28, HPA32, HPA35, HPA36, HPA34, HPA37, HPA8, HPA13, HPA15, HPA29, HPA33, the n terminal amino acid sequence of the expection of HPA38 and HPA41 is listed in SEQ ID No.1-8 respectively, and 20,21 and 25-31.

Utilize EMBL and GenBank (Release 91) and DNA STAR system that the cDNA sequence of determining of institute's isolated polypeptide and the aminoacid sequence of expection are compared with known sequences in the gene pool.DNA STAR system is the association of the protein sequence (Release 91) of Swiss, PIR database and translation.Do not find and HPA17 HPA25, HPA28, HPA32, the remarkable homologous of HPA35 and HPA36.

Find have an appointment 100% identity of cDNA sequence that HPA8 determines and people's proto-oncogene BMI-1 (Alkema, people such as M.J., human molecular genetics (Hum.Mol.Gen.) 2:1597-1603,1993).5 ＇ with coding HPA13 find and the known cDNA 100% identical (GenBank searching number D63880) that derives from people's prematurity myelocytic series with 3 ＇ cDNA sequence retrieval DNA databases.Infer that with HPA13 the aminoacid sequence retrieval Protein Data Bank discovery that is identical with the above-mentioned identical human cDNA sequence's of coding open reading frame 100%.Aminoacid sequence retrieval Protein Data Bank with the HPA15 expection, finding has high homology (60% is identical) (Swiss/PIR registration number S46677) with the open reading frame of yeast saccharomyces cerevisiae (Saccharomycescerevisiae) prediction, and with a kind of people's protein 10 0% identical (Lonnroth that derives from adjusting intestinal juice excretory pituitary gland, I., journal of biological chemistry (J.Bio.Chem.) 35:20615-20620,1995).The aminoacid sequence that discovery HPA38 infers is identical with human heat shock factor protein 2 (institute of NAS reports 88:6911-6915 for Schuetz, people such as T.J., 1991) 100%.Find and people LIM protein (Rearden with HPA415 ＇ dna sequence dna retrieval DNA database and with the aminoacid sequence retrieval Protein Data Bank of inferring, A., biological chemistry and biophysical studies communication (Biochem.Biophys.Res.Commun.) 201:1124-1131,1994) 100% is identical.Known to the limit inventor, remove LIM protein, be shown without any polypeptide of the present invention and be present in the human prostate.

Use the virion ehec infection XL-1 Blue MRF ＇ of positive phagemid, as described in people such as Sambrook, the same.Induce recombinant protein by adding IPTG.Induced with carry out the SDS-PAGE electrophoresis without the inductive cell pyrolysis liquid with identical form and transfer on the nitrocellulose filter.Filter membrane is respectively with the scorching serum of human prostate (dilution in 1: 200) with there is the active serum (1: 200 or dilution in 1: 250) of exempting from of LacZ N end 4Kd partial reaction to react.Serum was hatched 2 hours under the room temperature.By adding ¹²⁵The different time in the scope detected bonded antibody from 16 hours to 11 days to exposure behind the albumin A of I mark.Immuning hybridization the results are summarized in the table I, wherein (+) expression positive reaction, (-) expression is reactionless.

The scorching serum of table I antigen human prostate is anti--and lacZ serum proteins molecular weight/KdHPA8 (-) (-) HPA13 (+) (+) HPA15 (+) (+) 50HPA16 (+) (+) 40HPA17 (+) (-) 40HPA20 (+) (+) 38HPA25 (-) (+) 32HPA28 (-) (-) HPA29 (+) (+) HPA32 (-) (-) HPA33 (+) (+) HPA34 do not survey (+) 50HPA35 (-) (-) HPA36 (-) (-) HPA37 and do not survey (+) 50HPA38 (-) (-) HPA41 and do not survey (+)

Scorching serum of recombinant human prostatitis antigen and human prostate and anti-LacZ serum all have positive reaction to show that the reactivity of the scorching serum of human prostate is at this fusion rotein.Clone's antigen shows with the reactivity of the scorching serum of human prostate not and anti-LacZ sero-reaction, and it is inner to show that proteins C reactive may originate from the clone.Antigen and anti-LacZ sero-reaction but not with the scorching sero-reaction of human prostate may be in the scorching serum identification of human prostate conformational epitope's result or the Ag-Ab binding kinetics immuning hybridization expose 2 hours still not enough.Can't help escherichia coli expression with two kinds of all nonreactive antigens of serum, reactive epi-position can be within fusion rotein or inner open reading frame.Because from HPA13, the unstable of the recombinant antigen of HPA29 and HPA33 can't be determined the size of recombinant antigen.

With RT-PCR 4 kinds of different human cell lines (comprising that two metastatic prostate tumours are LNCaP and DU145), normal prostatic, breast, colon, kidney, stomach, lung and skeletal muscle tissue are detected the scorching antigenic expression of representative human prostate in 9 kinds of different tumor of prostate samples mammary tumor sample different with 3 kinds.Result of study sees Table II.

The analysis clone LNCaP DU145 MCF-HBL-prostate gland mammary gland colon kidney stomach lung skeletal muscle that the table II is expressed HPA clone mRNA in human cell line, healthy tissues and the tumour by the RT-PCR method

12A 100hpa-17 + ++ + + + - ± - - + +hpa-20 +++ ++++ NT NT ± NT NT - NT + NThpa-28 + +++ + + + - ± + - + ±

Tumor of prostate (n=9) tumor of breast (n=3) clone tumour 1 tumour 2 tumours 3 tumours 4 tumours 5 tumours 6 tumours 7 tumours 8 tumours 9 tumours 1 tumour 2 tumour 3hpa-17+++-++ ±--+++ ++ hpa-20++ NT NT NT NT NT NT NT++ +++hpa-28++ ±-++ ++ ±-++ ++++

Also detected representative antigenic mRNA expression in LNCaP and normal prostatic, kidney, liver, stomach, lung and pancreas by the RNase protection.Result of study sees Table III.

The branch that the table III is expressed HPA clone mRNA in LNCaP and the health adult tissue by the RNase protection

Analyse clone's LNCaP prostate kidney liver stomach lung pancreas hpa-15+-++ +++-++ hpa-20 ++ +++++++NT NThpa-25++++ ++ ++ NThpa-32 NT ++++ NT ++ NThpa-35 ++++++NT++ ++++ hpa-36++ NT NT+++Isolation and Identification of embodiment 2A. rat kind sterol conjugated protein

Immune serum is available from using the immunity of rat prostate extract to produce the rat at the antibody of self prostate antigen.Concrete, rat obtains control serum at the blood of looking ahead before with the immunity of the rat prostate stain remover extract (PBS that contains 0.1%Triton) in the Freund's complete adjuvant.Initial immunity carries out the incomplete Freund's adjuvant booster immunization after 3 weeks, and blood sampling is clear during 6 weeks.

With the scheme of manufacturers the serum of above-mentioned acquisition is carried out ECL Western hybridization analysis, confirmed a kind of rat prostate protein as shown in Figure 1.After the reduction, find the wide silver-colored colored zone of migration at the 7kD place through SDS-PAGE.Without reduction, at the visible strong band in 24kD place (Fig. 2).By this albumen of ion exchange chromatography purifying and the gel electrophoresis under reductive condition.As seen three bands, showing has three chains in the protein: 6-8kD chain (C1), 8-10kD chain (C2) and 10-12kD chain (C3).Utilize reversed-phase HPLC at Delta ^TM((Waters-Millipore, Milford are further purified this albumen on MA) to column dimension 3.9 * 300mm) to C18 300 5 μ m posts.Contain the proteic sample of 100 μ g and be dissolved in 0.1% trifluoroacetic acid (TFA), pH1.9, and polypeptide under acetonitrile linear gradient (0-60%) condition of 0.1%TFA (pH1.9) with the speed wash-out of 0.5ml/min 1 hour.Monitor elutriant at the 214nm place.Dimeric 2 peaks of C1-C3 dimer and C2-C3 have been obtained.The N-terminal of finding the C2 chain is closed.On the Procise Model of Perkin Elmer/AppliedBiosysterms company 494 protein sequencers, measure the sequence of C1 and C3 chain, find to have following N-terminal sequence (being respectively Seq ID No.44 and 45)

(a) Ser-Gln-Ile-Cys-Glu-Leu-Val-Ala-His-Glu-Thr-Ile-Ser-Phe-Leu; With

(b) Xaa-Xaa-Xaa-Xaa-Xaa-Ser-Ile-Leu-asp-Glu-Val-Ile-Arg-Gly-Thr, wherein Xaa can be any amino acid.

Utilize the database of discussing among the embodiment 1 that the known array in the homogenic storehouse of these sequences is compared, find and muroid sterol conjugated protein (being also referred to as estramustine conjugated protein (EMBP)) (Forsgren, B. wait the people, clinical biological study progress (Prog.Clin.Biol.Res.) 75A:391-407,1981; Forsgren, people such as B., institute of NAS reports 76:3149-53, and 1979) identical.This albumen is secretory protein main in the rat seminal fluid and demonstration and steroid, cholesterol and proline(Pro) have been rich in protein binding.EMBP has shown the active metabolite in conjunction with estramustine and estromustine, estramustine phosphate.Reported that estramustine phosphate is used for treating clinically the advanced prostate cancer (see example: Van Poppel, people such as H., clinical biological study progress 370:323-41,1991) that the standard hormone is broken away from the unresponsive patient of treatment.B. infer with rat homologous human steroid Isolation of Binding Proteins

From the rat prostate of fresh excision, obtain the steroid binding protein of purifying and be used for the pure female rabbit of subcutaneous immune New Zealand white that (the rat steroid binding protein of 150 μ g purifying contains 100 μ g Muramyl dipeptides (auxiliary peptide in 1mlPBS and 1ml, Calbiochem, La Jolla, Freund's incomplete adjuvant CA).After 6 weeks rabbit is used the same subcutaneous booster immunization of protein dosage in the Freund's incomplete adjuvant.At last, to the rabbit booster immunization, the back blood sampling of two week of immunity clearly the last time with the proteinic PBS intravenously of 100 μ g in 2 week backs.

With the failure of gained rabbit anti-serum screening Ln Cap.fgc clone.Collect liquid with this rabbit anti-serum screening with the anion-exchange chromatography that following embodiment 3 describes people's seminal fluid of scheme in detail subsequently.This analysis has disclosed a kind of about 18-22kD cross reactivity albumen.Purpose seminal fluid part (part 1) is separated into single component by SDS-PAGE under the irreducibility condition, hybridize on the pvdf membrane, cuts and digestion with CNBr in 70% formic acid.The CNBr fragment of gained is separated on Tris-glycine (tricine) gel systems, and electricity hybridizes to PVDF and is cut once more.The sequence of a peptide is determined as follows:

Val-Val-Lys-Thr-Tyr-Leu-Ile-Ser-Ser-Ile-Pro-Leu-Gln-Gly-Ala-Phe-Asn-Tyr-Lys-Tyr-Thr-Ala(SEQ.ID?No.46).

Utilize the database identical that these sequences are compared with known array in the gene pool with the front, unexpected discovery is identical with thick gallbladder disease liquid eggs white matter (gross cystic disease fluid protein), this protein has been found relevant (Murphy with metastatic breast cancer, L.C. wait the people, journal of biological chemistry (J.Biol.Chem.) 262:15236-15241,1987).Known to the inventor, this albumen is not found in male tissue before this.

As above-mentioned part 1 with steroid bonded ability with following method research.The rat steroid binding protein (RSBP) of purifying and part 1 are transferred on the nitrocellulose filter through behind the SDS-PAGE.Particularly, 1.5 μ g RSBP/ gel swimming lanes and 4 μ g parts, 1/ gel swimming lane are on the 4-20% gradient Laemmli glue (BioRad) abreast behind the electrophoresis, and electrophoretic transfer is to nitrocellulose filter again.After protein shifts, in the PBS of 1%Tween20, sealed 1 hour under the filter membrane room temperature, each with containing the PBS10ml washing 3 times that 0.1%Tween20 adds 0.5M NaCl in addition, each 10 minutes, using 1 again) 0.87 μ M is coupled to the progesterone in the washings of being diluted in of horseradish peroxidase (HRP); 2) 0.87 μ M progesterone HRP and 200 μ M estramustines; Or 3) 0.87 μ M progesterone HRP adds that the estramustine of unlabelled progesterone of 400 μ M and 200 μ M detects.Each reaction mixture was all at room temperature hatched 1 hour and was used 0.1%Tween20, PBS and 0.5M NaCl washing 3 times, each 10 minutes.(ECL systerm Amersham) also can be conjugated protein in conjunction with the progesterone HRP of estramustine to show to the hybridization colour developing.

For rat steroid binding protein and part 1, all obtain three bands, promptly compete bonded band (Fig.3) in conjunction with the band of HRP-progesterone with unlabelled progesterone and estramustine.These results show from as above-mentioned people's seminal fluid isolating three bands in conjunction with hormone and on the quantity of polypeptide corresponding to the protein-bonded C1 of rat kind sterol, C2 and C3 chain, though or since the modification after the translation of primary sequence or secondary make its molecular weight more bigger.

The protein-bonded homologue of rat kind sterol of this supposition is also found when screening people's seminal fluid with above-mentioned rabbit anti-serum subsequently.Obtained a kind of hydrophobicity 22kD/65kD particularly and assembled protein, behind CNBr digestion 22kD band, it provides the peptide with following sequence:

Val-Val-Lys-Thr-Tyr-Leu-Ile-Ser-Ser-Ile-Pro-Leu-Gln-Ala-Phe-Asn-Tyr-Lys-Tyr-Thr-Ala (SEQ.ID No.47). find that this peptide is corresponding with proteic the 67th residue to the of thick gallbladder disease liquid 87 residues, and utilize human autoimmune prostatitis serum to identify once more, such as following embodiment 4 discussion.Embodiment 3 utilizes the separation and the evaluation of the scorching serum of rat prostate isolated polypeptide from Ln Cap.fgc

By Ln Cap.fgc cell precipitation is resuspended in PBS, 1%NP-40 and 60 μ g/ml phenylmethylsulfonyl fluoride (PMSF) (Sigma, St.Louis, MO) back in the Dounce refiner after handling for 10 times by homogenization (10gm cell precipitation/10ml).Be 30 seconds probe supersound process subsequently and in the Dounce refiner, handle through 10 times again.Gained homogenate is centrifugal with the speed of 10,000 * G, and (Amicon, Beverly are used for BioRad PrepQ-20 anionite-exchange resin after MA) to supernatant liquor with 0.45 μ m membrane filtration.Protein 20mM tris, among the pH7.5 in 70 minutes 0 to the NaCl gradient of 0.8M speed wash-out with 8ml/min.Collect concentrated and-20 ℃ of storages under the condition that has 60 μ g/ml PMSF to exist of component cooling back with centrifugal type (centriprep) concentrating instrument (Amicon) of 10kD MWCO.Ion-exchange is collected and is transferred on the nitrocellulose filter after liquid is used 4-20%tris glycine Ready-Gels (BioRad) electrophoresis detection subsequently.Utilize the rat blood serum among the foregoing description 3A to collect liquid by the exchange of ECL (Amersham International) Western Analysis and Identification object ion.The protein of a kind of about 65kD of this analysis revealed arrives 0.13MNaCl place wash-out 0.08.Having the reactive ion-exchange of rat blood serum to collect liquid successively analyzes to identify the target protein part with HPLC and Western.This protein is again in order to digestion in saturated 70% formic acid of the CNBr of methionine residues place cutting 24 hours.

Gained CNBr fragment is with micropore HPLC purifying, used be Vydac C18 post (Hesperia, CA), column dimension 1 * 150mm, Perkin Elmer/Applied Biosysterms company (Foster City, Division Model 172 HPLC CA).With the 0-60% acetonitrile gradient with the speed of 40 μ l/min elution samples from the post.At 214nm place monitoring elutriant.Gained level part is directly gone up to the Perkin Elmer/Applied Biosysterms Procise of company 494 protein sequencers and is checked order from aminoterminal with the Edman chemical method of standard.Obtained two different peptides with following sequence:

(a) Xaa-Ala-Lys-Lys-Phe-Leu-Asp-Ala-Glu-His-Lys-Leu-Asn-Phe-Ala (SEQ.ID No.48); With

(b) Xaa-Xaa-Xaa-Lys-Ile-Lys-Lys-Phe-Ile-Gln-Glu-Asn-Ile-Phe-Gly, wherein Xaa can be any amino acid (SEQ ID No.49)

Utilize the same database in front that these sequences are compared with known sequences in the gene pool, be accredited as the 286th residue to the 300 residues and the 228th residue to the 242 residues of possible protein disulfide isomerase ER-60 precursor respectively, this precursor is called ER-60 (Bado herein, R.J. wait the people, incretology (Endocrinology) 123:1264-1273,1988).This antigen also is known as Phospholipase C-α (seeing PCT WO 95/08624).285 and 227 residues of ER-60 are methionine(Met), and are consistent with above-mentioned cyanogen bromide fragments sequence.

ER-60 has the active resident endoplasm albumen of various biological, comprises disulfide bond isomerase and restricted cysteine protease activity.Concrete, ER-60 has shown preferential degraded calnexin, and the latter is the protein of a kind of participation by the angtigen presentation of the main histocompatibility complex (MHC) of I class (Class I) approach.ER-60 has been found in overexpression in the colorectal carcinoma with relevant family member ER-72, and the ER-60 of clipped form shows enhanced the enzyme activity (Egea, people such as G., cell science magazine (J.Cell.Sci.) (Britain) 105:819-30,1993).Yet just known to the inventor, this peptide species does not still know to occur or overexpression in human prostate before this.Recently, ER-60 genetic expression is by be associated with the contact inhibition of inducing cell propagation (Greene, people such as J.J., cellular elements biology (Cell.Mol.Biol.) 41:473-80,1995).Therefore, if ER-60 is also functional by brachymemma and right and wrong in prostate cancer, in the colorectal carcinoma, the product that lacks contact inhibition will cause neoplastic transformation and tumor development as it.Embodiment 4 utilizes the separation and the evaluation of the scorching serum of human prostate isolated polypeptide from Ln CaP.fgc

Utilization as ion exchange technique as described in the embodiment 3 with the scorching serum screening Ln of human prostate described in the embodiment 1 CaP.fgc clone.Collect liquid with reversed-phase HPLC purification reaction ion-exchange as the aforementioned, utilize with described sero-fast cross reaction as choice criteria, separated the polypeptide that is shown in SEQ IDNo.50-51.With above-mentioned database relatively finding with known array in these sequences and the gene pool as showing the homologue shown in the II.Yet any of these polypeptide never is associated with human prostate before this.

Table IV SEQ ID No. database retrieval identifies that 53 glyceraldehyde-3-phosphate dehydrogenases, 54 α-white embodiment 5 of people's fructose-bis phosphate aldolase 55 calprotectins 56 calprotectins white 59 gallbladder disease liquid eggs of 57 malate dehydrogenase (malic acid dehydrogenase)s, 58 gallbladder disease liquid eggs is derived from the separation and the evaluation of the polypeptide of people's seminal fluid

The polypeptide that is derived from people's seminal fluid by the anion-exchange chromatography purifying reaches homogeneity.Particularly, with 0.1mM Bis-Tris propane damping fluid (pH7) with upper prop behind 10 times of the seminal fluid diluted samples.Utilize gel perfusion chromatography (gel profusion chromatography) by the last fraction collection polypeptide of 0.01mM Bis-Tris propane damping fluid (pH7.5) equilibrated Poros (Perseptive Biosysterms) 146 II Q/M anion-exchange column 4.6mm * 10mm.Protein is with 0～0.5MNaCl linear gradient elution in the above-mentioned damping fluid.At 220nm wavelength place monitoring post elutriant.Each stream part further by reversed-phase HPLC at Vydac (Hesperia, CA) purifying on the C18 post.

Measure the sequence of gained part as above-mentioned embodiment 3.Obtained a peptide with following N-terminal sequence:

(c)Met-Asp-Ile-Pro-Gln-Thr-Lys-Gln-Asp-Leu-Glu-Leu-Pro-Lys-Leu

(SEQ ID NO:57). as above-mentioned with known sequences in these sequences and those gene pools compare find identical with people's placental protein 14 (PP14) 100%.Synthesizing of embodiment 6 polypeptide

Utilization is by HPTU (0-benzotriazole-N, N, N ＇, N ＇-tetramethyl-urine hexafluorophosphate) activatory FMOC chemical method synthetic polypeptide on Applied Biosysterms 430A peptide synthesizer.Can add that a Gly-Cys-Gly sequence is to provide coupling, to be attached to the method for immobilized surface or this peptide of mark at the aminoterminal of this peptide.Can this peptide be cut down from solid support with following cutting mixture: trifluoroacetic acid: dithioglycol: thioanisole: water: phenol (40: 1: 2: 2: 3).Cut after 2 hours, this peptide can precipitate with cold methyl tertiary butyl ether.Precipitation is again with after water dissolution that contains 0.1% trifluoroacetic acid (TFA) and cold the doing, with C18 reversed-phase HPLC purifying.Can utilize this peptide of 0%-60% acetonitrile (containing 0.1%TFA) gradient elution that (contains 0.1%TFA) in the water.After cold dried pure level part, utilize the mass spectrum of electrospray mass spectrum or other type and the characteristic that amino acid analysis is determined this peptide.

Though from preamble narration, should be appreciated that and described specific embodiments of the present invention herein for purposes of illustration multiple modification to be arranged and without departing from the spirit and scope of the present invention.

Sequence table (1) general information

(ⅰ) applicant: Corixa company

(ⅱ) invention exercise question: the compound and the method that are used for the immunotherapy and the immunodiagnosis of prostate cancer

(ⅲ) sequence number: 57

(ⅳ) address:

(A) address: SEED and BERRY LLP

(B) street: 6300 Columbia Center, 701 Fifth Avenue

(C) city: Seattle

(D) state: Washington

(E) country: the U.S.

(F) postcode code: 98104-7092

(ⅴ) computer-reader form:

(A) media type: floppy disk

(B) computer: IBM PC compatible

(C) operating system: PC-DOS/MS-DOS

(D) software: Patent In Release#1.0, Version# 1.30

(ⅵ) current request for data

(A) application number:

(B) Date to Tender Notice of Readiness: 1997-5-14

(C) classification:

(ⅷ) lawyer/proxy's information:

(A) name: Maki, David J.

(B) number of registration: 31,392

(C) reference/reel number: 210121.424PC

(ⅸ) communication information:

(A) phone: (206) 622-4900

(B) fax: the information of (206) 682-6031 (2) SEQ ID No.1

(ⅰ) sequence signature:

(A) length: 89 amino acid

(B) type: amino acid

(C) chain:

(D) topological classification: the sequence description of linear (ⅹ ⅰ) SEQ ID NO:1: Ala Arg Ala Ser Val Met Leu Leu Gly Met Met Ala Arg Gly Lys Pro1 5 10 15Glu Ile Val Gly Ser Asn Leu Asp Thr Leu Met Ser Ile Gly Leu Asp

20 25 30Glu?Lys?Phe?Pro?Gln?Asp?Tyr?Arg?Leu?Ala?Gln?Gln?Val?Cys?His?Ala

35 40 45Ile?Ala?Asn?Ile?Ser?Asp?Arg?Arg?Lys?Pro?Ser?Leu?Gly?Lys?Arg?His

50 55 60Pro?Pro?Phe?Arg?Leu?Pro?Gln?Glu?His?Arg?Leu?Phe?Glu?Arg?Leu?Arg65 70 75 80Glu?Thr?Val?Thr?Lys?Gly?Phe?Val?His

The information of 85 (2) SEQ ID No.2

(ⅰ) sequence signature:

(A) length: 89 amino acid

(B) type: amino acid

(C) chain:

(D) topological classification: the sequence description of linear (ⅹ ⅰ) SEQ ID NO:2: Ala Arg Gly Arg Phe Gly Arg Leu Gly Val Gly Gly Glu Pro His Pro1 5 10 15Arg Arg Asn Pro Ala Leu Pro Thr Glu Leu Ala Glu Leu Thr Pro Gln

20 25 30Val?Arg?Arg?Ala?Ala?Xaa?Lys?Thr?Gln?Arg?Ser?Gln?Val?Lys?Pro?Arg

35 40 45His?Arg?Arg?Gly?Trp?Pro?Pro?Thr?Val?Pro?Leu?Ala?Gly?Arg?Leu?Glu

50 55 60Glu?Leu?Lys?Thr?Pro?Arg?Ser?Pro?Arg?Pro?Pro?Glu?Gln?Gly?Leu?Asp65 70 75 80Pro?Ser?Pro?Cys?Ser?Leu?Pro?Ser?Pro

The information of 85 (2) SEQ ID No.3

(ⅰ) sequence signature:

(A) length: 858 amino acid

(B) type: amino acid

(C) chain:

(D) topological classification: the sequence description of linear (ⅹ ⅰ) SEQ ID NO:3: Gln Glu Ser Glu Pro Phe Ser His Ile Asp Pro Glu Glu Ser Glu Glu1 5 10 15Thr Arg Leu Leu Asn Ile Leu Gly Leu Ile Phe Lys Gly Pro Ala Ala

20 25 30Ser?Thr?Gln?Glu?Lys?Asn?Pro?Arg?Glu?Ser?Thr?Gly?Asn?Met?Val?Thr

35 40 45Gly?Gln?Thr?Val?Cys?Lys?Asn?Lys?Pro?Asn?Met?Ser?Asp?Pro?Glu?Glu

50 55 60Ser?Arg?Gly?Asn?Asp?Glu?Leu?Val?Lys?Gln?Glu?Met?Leu?Val?Gln?Tyr65 70 75 80Leu?Gln?Asp?Ala?Tyr?Ser?Phe?Ser?Arg?Lys?Ile?Thr?Glu?Ala?Ile?Gly

85 90 95Ile?Ile?Ser?Lys?Met?Met?Tyr?Glu?Asn?Thr?Thr?Thr?Val?Val?Gln?Glu

100 105 110Val?Ile?Glu?Xaa?Phe?Val?Met?Val?Phe?Gln?Phe?Gly?Val?Pro?Gln?Ala

115 120 125Leu?Phe?Gly?Val?Arg?Arg?Met?Leu?Pro?Leu?Ile?Trp?Ser?Lys?Glu?Pro

130 135 140Gly?Val?Arg?Glu?Ala?Val?Leu?Asn?Ala?Tyr?Arg?Gln?Leu?Tyr?Leu?Asn145 150 155 160Pro?Lys?Gly?Asp?Ser?Ala?Arg?Ala?Lys?Ala?Gln?Ala?Leu?Ile?Gln?Asn

165 170 175Leu?Ser?Leu?Leu?Leu?Val?Asp?Ala?Ser?Val?Gly?Thr?Ile?Gln?Cys?Leu

180 185 190Glu?Glu?Ile?Leu?Cys?Glu?Phe?Val?Gln?Lys?Asp?Glu?Leu?Lys?Pro?Ala

195 200 205Val?Thr?His?Leu?Leu?Trp?Glu?Arg?Ala?Thr?Glu?Lys?Val?Ala?Cys?Cys

210 215 220Pro?Leu?Glu?Arg?Cys?Ser?Ser?Val?Met?Leu?Leu?Gly?Met?Met?Ala?Arg225 230 235 240Arg?Lys?Pro?Glu?Ile?Val?Gly?Ser?Asn?Leu?Asp?Thr?Leu?Met?Ser?Ile

245 250 255Gly?Leu?Asp?Glu?Lys?Phe?Pro?Gln?Asp?Tyr?Arg?Leu?Ala?Gln?Gln?Val

260 265 270Cys?His?Ala?Ile?Ala?Asn?Ile?Ser?Asp?Arg?Arg?Lys?Pro?Ser?Leu?Gly

275 280 285Lys?Arg?His?Pro?Pro?Phe?Arg?Leu?Pro?Gln?Glu?His?Arg?Leu?Phe?Glu

290 295 300Arg?Leu?Arg?Glu?Thr?Val?Thr?Lys?Gly?Phe?Val?His?Pro?Asp?Pro?Leu305 310 315 320Trp?Ile?Pro?Phe?Lys?Glu?Val?Ala?Val?Thr?Leu?Ile?Tyr?Gln?Leu?Ala

325 330 335Glu?Gly?Pro?Glu?Val?Ile?Cys?Ala?Gln?Ile?Leu?Gln?Gly?Cys?Ala?Lys

340 345 350Gln?Ala?Leu?Glu?Lys?Leu?Glu?Glu?Lys?Arg?Thr?Ser?Gln?Glu?Asp?Pro

355 360 365Lys?Glu?Ser?Pro?Ala?Met?Leu?Pro?Thr?Phe?Leu?Leu?Met?Asn?Leu?Leu

370 375 380Ser?Leu?Ala?Gly?Asp?Val?Ala?Leu?Gln?Gln?Leu?Val?His?Leu?Glu?Gln385 390 395 400Ala?Val?Ser?Gly?Glu?Leu?Cys?Arg?Arg?Arg?Val?Leu?Arg?Glu?Glu?Gln

405 410 415Glu?His?Lys?Thr?Lys?Asp?Pro?Lys?Glu?Lys?Asn?Thr?Ser?Ser?Glu?Thr

420 425 430Thr?Met?Glu?Glu?Glu?Leu?Gly?Leu?Val?Gly?Ala?Thr?Ala?Asp?Asp?Thr

435 440 445Glu?Ala?Glu?Leu?Ile?Arg?Gly?Ile?Cys?Glu?Met?Glu?Leu?Leu?Asp?Gly

450 455 460Lys?Gln?Thr?Leu?Ala?Ala?Phe?Val?Pro?Leu?Leu?Leu?Lys?Val?Cys?Asn465 470 475 480Asn?Pro?Gly?Leu?Tyr?Ser?Asn?Pro?Asp?Leu?Ser?Ala?Ala?Ala?Ser?Leu

485 490 495Ala?Leu?Gly?Lys?Phe?Cys?Met?Ile?Ser?Ala?Thr?Phe?Cys?Asp?Ser?Gln

500 505 510Leu?Arg?Leu?Leu?Phe?Thr?Met?Leu?Glu?Lys?Ser?Pro?Leu?Pro?Ile?Val

515 520 525Arg?Ser?Asn?Leu?Met?Val?Ala?Thr?Gly?Asp?Leu?Ala?Ile?Arg?Phe?Pro

530 535 540Asn?Leu?Val?Asp?Pro?Trp?Thr?Pro?His?Leu?Tyr?Ala?Arg?Leu?Arg?Asp545 550 555 560Pro?Ala?Gln?Gln?Val?Arg?Lys?Thr?Ala?Gly?Leu?Val?Met?Thr?His?Leu

565 570 575Ile?Leu?Lys?Asp?Met?Val?Lys?Val?Lys?Gly?Gln?Val?Ser?Glu?Met?Ala

580 585 590Val?Leu?Leu?Ile?Asp?Pro?Glu?Pro?Gln?Ile?Ala?Ala?Leu?Ala?Lys?Asn

595 600 605Phe?Phe?Asn?Glu?Leu?Ser?His?Lys?Gly?Asn?Ala?Ile?Tyr?Asn?Leu?Leu

610 615 620Pro?Asp?Ile?Ile?Ser?Arg?Leu?Ser?Asp?Pro?Glu?Leu?Gly?Val?Glu?Glu625 630 635 640Glu?Pro?Phe?His?Thr?Ile?Met?Lys?Gln?Leu?Leu?Ser?Tyr?Ile?Thr?Lys

645 650 655Asp?Lys?Gln?Thr?Glu?Ser?Leu?Val?Glu?Lys?Leu?Cys?Gln?Arg?Phe?Arg

660 665 670Thr?Ser?Arg?Thr?Glu?Arg?Gln?Gln?Arg?Asp?Leu?Ala?Tyr?Cys?Val?Ser

675 680 685Gln?Leu?Pro?Leu?Thr?Glu?Arg?Gly?Leu?Arg?Lys?Met?Leu?Asp?Asn?Phe

690 695 700Asp?Cys?Phe?Gly?Asp?Lys?Leu?Ser?Asp?Glu?Ser?Ile?Phe?Ser?Ala?Phe705 710 715 720Leu?Ser?Val?Val?Gly?Lys?Leu?Arg?Arg?Gly?Ala?Lys?Pro?Glu?Gly?Lys

725 730 735Ala?Ile?Ile?Asp?Glu?Phe?Glu?Gln?Lys?Leu?Arg?Ala?Cys?His?Thr?Arg

740 745 750Gly?Leu?Asp?Gly?Ile?Lys?Glu?Leu?Glu?Ile?Gly?Gln?Ala?Gly?Ser?Gln

755 760 765Arg?Ala?Pro?Ser?Ala?Lys?Lys?Pro?Ser?Thr?Gly?Ser?Arg?Tyr?Gln?Pro

770 775 780Leu?Ala?Ser?Thr?Ala?Ser?Asp?Asn?Asp?Phe?Val?Thr?Pro?Glu?Pro?Arg785 790 795 800Arg?Thr?Thr?Arg?Arg?His?Pro?Asn?Thr?Gln?Gln?Arg?Ala?Ser?Lys?Lys

805 810 815Lys?Pro?Lys?Val?Val?Phe?Ser?Ser?Asp?Glu?Ser?Ser?Glu?Glu?Asp?Leu

820 825 830Ser?Ala?Glu?Met?Thr?Glu?Asp?Glu?Thr?Pro?Lys?Lys?Thr?Thr?Pro?Ile

835 840 845Leu?Arg?Ala?Ser?Ala?Arg?Arg?His?Arg?Ser

The information of 850 855 (2) SEQ ID No.4

(ⅰ) sequence signature:

(A) length: 127 amino acid

(B) type: amino acid

(C) chain:

(D) topological classification: the sequence description of linear (ⅹ ⅰ) SEQ ID NO:4: Ala Arg Asp Arg Leu Val Ala Ser Lys Thr Asp Gly Lys Ile Val Gln1 5 10 15Tyr Glu Cys Glu Gly Asp Thr Cys Gln Glu Glu Lys Ile Asp Ala Leu

20 25 30Gln?Leu?Glu?Tyr?Ser?Tyr?Leu?Leu?Thr?Ser?Gln?Leu?Glu?Ser?Gln?Arg

35 40 45Ile?Tyr?Trp?Glu?Asn?Lys?Ile?Val?Arg?Ile?Glu?Lys?Asp?Thr?Ala?Glu

50 55 60Glu?Ile?Asn?Asn?Met?Lys?Thr?Lys?Phe?Lys?Glu?Thr?Ile?Xaa?Xaa?Cys65 70 75 80Asp?Asn?Leu?Glu?His?Xaa?Leu?Asn?Asp?Leu?Leu?Lys?Glu?Lys?Gln?Ser

85 90 95Val?Glu?Arg?Lys?Cys?Thr?Gln?Leu?Asn?Thr?Lys?Val?Ala?Lys?Leu?Thr

100 105 110Asn?Glu?Leu?Lys?Glu?Glu?Gln?Glu?Met?Asn?Lys?Cys?Leu?Arg?Ala

The information of 115 120 125 (2) SEQ ID No.5

(ⅰ) sequence signature:

(A) length: 43 amino acid

(B) type: amino acid

(C) chain:

(D) topological classification: the sequence description of linear (ⅹ ⅰ) SEQ ID NO:5: Ala Arg Ala Glu Val Gln Arg Trp Arg Arg Leu Val Ala Gly Arg Arg1 5 10 15Arg Ala Gly Gly Asp Gly Gly Asn Ser Gly Ser Cys Ser Arg Trp Gly

20 25 30Gly?Phe?Thr?Ser?Tyr?Pro?Trp?Asp?Arg?Glu?Ile

The information of 35 40 (2) SEQ ID No.6

(ⅰ) sequence signature:

(A) length: 751 amino acid

(B) type: amino acid

(C) chain:

(D) topological classification: the sequence description of linear (ⅹ ⅰ) SEQ ID NO:6: Pro Ala Glu Ala His Ser Asp Ser Leu Ile Asp Thr Phe Pro Glu Cys1 5 10 15Ser Thr Glu Gly Phe Ser Ser Asp Ser Asp Leu Val Ser Leu Thr Val

20 25 30Asp?Val?Asp?Ser?Leu?Ala?Glu?Leu?Asp?Asp?Gly?Met?Ala?Ser?Asn?Gln

35 40 45Asn?Ser?Pro?Ile?Arg?Thr?Phe?Gly?Leu?Asn?Leu?Ser?Ser?Asp?Ser?Ser

50 55 60Ala?Leu?Gly?Ala?Val?Ala?Ser?Asp?Ser?Glu?Gln?Ser?Lys?Thr?Glu?Glu65 70 75 80Glu?Arg?Glu?Ser?Arg?Ser?Leu?Phe?Pro?Gly?Ser?Leu?Lys?Pro?Lys?Leu

85 90 95Gly?Lys?Arg?Asp?Tyr?Leu?Glu?Lys?Ala?Gly?Glu?Leu?Ile?Lys?Leu?Ala

100 105 110Leu?Lys?Lys?Glu?Glu?Glu?Asp?Asp?Tyr?Glu?Ala?Ala?Ser?Asp?Phe?Tyr

115 120 125Arg?Lys?Gly?Val?Asp?Leu?Leu?Leu?Glu?Gly?Val?Gln?Gly?Glu?Ser?Ser

130 135 140Pro?Thr?Arg?Arg?Glu?Ala?Val?Lys?Arg?Arg?Thr?Ala?Glu?Tyr?Leu?Met145 150 155 160Arg?Ala?Glu?Ser?Ile?Ser?Ser?Leu?Tyr?Gly?Lys?Pro?Gln?Leu?Asp?Asp

165 170 175Val?Ser?Gln?Pro?Pro?Gly?Ser?Leu?Ser?Ser?Arg?Pro?Leu?Trp?Asn?Leu

180 185 190Arg?Ser?Pro?Ala?Glu?Glu?Leu?Lys?Ala?Phe?Arg?Val?Leu?Gly?Val?Ile

195 200 205Asp?Lys?Val?Leu?Leu?Val?Met?Asp?Thr?Arg?Thr?Glu?His?Thr?Phe?Ile

210 215 220Leu?Xaa?Gly?Leu?Arg?Lys?Ser?Ser?Glu?Tyr?Ser?Arg?Asn?Arg?Lys?Thr225 230 235 240Ile?Xaa?Pro?Arg?Cys?Val?Pro?Xaa?Met?Val?Cys?Leu?His?Lys?Tyr?Ile

245 250 255Ile?Ser?Glu?Glu?Ser?Xaa?Phe?Leu?Val?Leu?Gln?His?Ala?Glu?Xaa?Gly

260 265 270Lys?Leu?Trp?Ser?Tyr?Ile?Ser?Lys?Phe?Leu?Asn?Arg?Ser?Pro?Glu?Glu

275 280 285Ser?Phe?Asp?Ile?Lys?Glu?Val?Lys?Lys?Pro?Thr?Leu?Ala?Lys?Val?His

290 295 300Leu?Gln?Gln?Pro?Thr?Ser?Ser?Pro?Gln?Asp?Ser?Ser?Ser?Phe?Glu?Ser305 310 315 320Arg?Gly?Ser?Asp?Gly?Gly?Ser?Met?Leu?Lys?Ala?Leu?Pro?Leu?Lys?Ser

325 330 335Ser?Leu?Thr?Pro?Ser?Ser?Gln?Asp?Asp?Ser?Asn?Gln?Glu?Asp?Asp?Gly

340 345 350Gln?Asp?Ser?Ser?Pro?Lys?Trp?Pro?Asp?Ser?Gly?Ser?Ser?Ser?Glu?Glu

355 360 365Glu?Cys?Thr?Thr?Ser?Tyr?Leu?Thr?Leu?Cys?Asn?Glu?Tyr?Gly?Gln?Glu

370 375 380Lys?Ile?Glu?Pro?Gly?Ser?Leu?Asn?Glu?Glu?Pro?Phe?Met?Lys?Thr?Glu385 390 395 400Gly?Asn?Gly?Val?Asp?Thr?Lys?Ala?Ile?Lys?Ser?Phe?Pro?Ala?His?Leu

405 410 415Ala?Ala?Asp?Ser?Asp?Ser?Pro?Ser?Thr?Gln?Leu?Arg?Ala?His?Glu?Leu

420 425 430Lys?Phe?Phe?Pro?Asn?Asp?Asp?Pro?Glu?Ala?Val?Ser?Ser?Pro?Arg?Thr

435 440 445Ser?Asp?Ser?Leu?Ser?Arg?Ser?Lys?Asn?Ser?Pro?Met?Glu?Phe?Phe?Arg

450 455 460Ile?Asp?Ser?Lys?Asp?Ser?Ala?Ser?Glu?Leu?Leu?Gly?Leu?Asp?Phe?Gly465 470 475 480Glu?Lys?Leu?Tyr?Ser?Leu?Lys?Ser?Glu?Pro?Leu?Lys?Pro?Phe?Phe?Thr

485 490 495Leu?Pro?Asp?Gly?Asp?Ser?Ala?Ser?Arg?Ser?Phe?Asn?Thr?Ser?Glu?Ser

500 505 510Lys?Val?Glu?Phe?Lys?Ala?Gln?Asp?Thr?Ile?Ser?Arg?Gly?Ser?Asp?Asp

515 520 525Ser?Val?Pro?Val?Ile?Ser?Phe?Lys?Asp?Ala?Ala?Phe?Asp?Asp?Val?Ser

530 535 540Gly?Thr?Asp?Glu?Gly?Arg?Pro?Asp?Leu?Leu?Val?Asn?Leu?Pro?Gly?Glu545 550 555 560Leu?Glu?Ser?Thr?Arg?Glu?Ala?Ala?Ala?Met?Gly?Pro?Thr?Lys?Phe?Thr

565 570 575Gln?Thr?Asn?Ile?Gly?Ile?Ile?Glu?Asn?Lys?Leu?Leu?Glu?Ala?Pro?Asp

580 585 590Val?Leu?Cys?Leu?Arg?Leu?Ser?Thr?Glu?Gln?Cys?Gln?Ala?His?Glu?Glu

595 600 605Lys?Gly?Ile?Glu?Glu?Leu?Ser?Asp?Pro?Set?Gly?Pro?Lys?Set?Tyr?Ser

610 615 620Ile?Thr?Glu?Lys?His?Tyr?Ala?Gln?Glu?Asp?Pro?Arg?Met?Leu?Phe?Val625 630 635 640Ala?Xaa?Val?Asp?His?Ser?Ser?Ser?Gly?Asp?Met?Ser?Leu?Leu?Pro?Ser

645 650 655Ser?Asp?Pro?Lys?Phe?Gln?Gly?Leu?Gly?Val?Val?Glu?Ser?Xaa?Val?Thr

660 665 670Ala?Asn?Asn?Thr?Glu?Glu?Ser?Leu?Phe?Arg?Ile?Cys?Ser?Pro?Leu?Ser

675 680 685Gly?Ala?Asn?Glu?Tyr?Ile?Ala?Ser?Thr?Asp?Thr?Leu?Lys?Thr?Glu?Glu

690 695 700Val?Leu?Leu?Phe?Thr?Asp?Gln?Thr?Asp?Asp?Leu?Ala?Lys?Glu?Glu?Pro705 710 715 720Thr?Ser?Leu?Phe?Xaa?Arg?Asp?Ser?Glu?Thr?Lys?Gly?Glu?Ser?Gly?Leu

725 730 735Val?Leu?Glu?Gly?Asp?Lys?Glu?Ile?His?Gln?Ile?Phe?Glu?Gly?Pro

The information of 740 745 750 (2) SEQ ID No.7

(ⅰ) sequence signature:

(A) length: six amino acid

(B) type: amino acid

(C) chain:

(D) topological classification: the sequence description of linear (ⅹ ⅰ) SEQ ID NO:7: the information of Ala Arg Gly Ser Thr Gln1 5 (2) SEQ ID No.8

(ⅰ) sequence signature:

(A) length: 16 amino acid

(B) type: amino acid

(C) chain:

(D) topological classification: the sequence description of linear (ⅹ ⅰ) SEQ ID NO:8: the information of Ala Arg Gly Ser Ser Gln Val Arg Val Lys Ser Trp Arg Gly Asp Met1 5 10 15 (2) SEQ ID No.9

(ⅰ) sequence signature:

(A) length: 271 base pairs

(B) type: nucleic acid

(C) chain: strand

(D) topological classification: the sequence description of linear (ⅹ ⅰ) SEQ ID NO:9: the information of CCGCACGAGC CTCTGTCATG CTTCTTGGCA TGATGGCACG AGGAAAGCCA GAAATTGTGG 60GAAGCAATTT AGACACACTG ATGAGCATAG GGCTGGATGA GAAGTTTCCA CAGGACTACA 120GGCTGGCCCA GCAGGTGTGC CATGCCATTG CCAACATCTC GGACAGGAGA AAGCCTTCTC 180TGGGCAAACG TCACCCCCCC TTCCGGCTGC CTCAGGAACA CAGGTTGTTT GAGCGACTGC 240GGGAGACAGT CACAAAAGGC TTTGTCCACC C 271 (2) SEQ ID No.10

(ⅰ) sequence signature:

(A) length: 403 base pairs

(B) type: nucleic acid

(C) chain: strand

(D) topological classification: the sequence description of linear (ⅹ ⅰ) SEQ ID NO:10: the information of GGGTGGATAA CCTGAGGTAG GGAGTTCGAG ACCAGCCTGA CCAACATGGA GAAACCCCAT 60CTCTACTAAA AATAAAAAAT TAGCCGGCGT ATTGGCGTGC GCCTGTAATC CCAGCTACTC 120AAGAGGCTGA GGCAGGAGAA TCGCCTGAAC CCAGAGGCGG AGGTTGTAGT GAGCCGAAAT 180CACACCATTG CACTCCAGCT TGGGCAACAA TAGCGAACCT CCATCTCAAA TTAAAAAAAA 240AATGCCTACA CGCTTCTTTA AAATGCAAGG CTTTCTCTTA AATTAGCCTA ACTGAACTGC 300GTTGAGCTGC TTCAACTTTG GAATATATGT TTGCCAATCT CCTTGTTTTC TAATGAATAA 360ATGTTTTTAT ATACTTTTAA AAAAAAAAAA AAAAAAACTC GAG 403 (2) SEQ ID No.11

(ⅰ) sequence signature:

(A) length: 2276 base pairs

(B) type: nucleic acid

(C) chain: strand

( D ) ： ( ⅹⅰ ) SEQ ID NO:11：GGAGGTTTGG GCGGCTTGGC GTCGGAGGAG AGCCCCACCC GCGGAGGAAC CCAGCCTTGC 60CAACGGAGCT GGCGGAGCTC ACTCCTCAGG TCAGGCGGGC GGCGTANAAA ACGCAGCGGA 120GCCAGGTGAA ACCAAGGCAC CGCCGTGGCT GGCCCCCGAC AGTTCCTCTA GCCGGGAGGT 180TGGAGGAGCT GAAAACGCCG CGGAGCCCTC GGCCGCCCGA GCAGGGGCTG GACCCCAGCC 240CTTGCAGCCT CCCTTCTCCT GGCACCCAAG TGCAGTCCTG GCTGCAGAAG GGGCCGCGGG 300CGCACTGAGT TTCCAACCTC CGTTCAGCCT GTCTGTCTCA GGGTGCAGCC TTAATGAGAG 360GTGATTCCTA AGCTGCTGGG AACCTGAGGT TGTCAAAGGG GCGGCAGGAA ATGGACAGCA 420GTATAAAACC CAGAAGCAGA ACTTGAAGGT TAAACCACTA GCCCATTTCA CAGAATGTTT 480CATCCATTTG TGGACCAAAA GATGGAGTTG GTTTTTATTT TTAAAAAGAT AATGTTAATG 540ATCTGATACC ACTACAAATA TTTACGTGAG AAGATTCATG GACTTGTCTT TTGGTTGGAC 600TGTCACTCAT TTCTGAAAGT TTCTTCAGCC ACAATTTCTA TTTGAAAATT CAAGTATCAA 660AGGATACCAG GTTTAGAATG GTATAATGAT GTATTTTGTC TGAGGACTGC AAATTTTATA 720GAGACCACAG TTGGATTCCA GTGATATTCT GCAATCAAAG TGATTTGATA AACCTAATTT 780TGAAGCATTT TATATTTATA AGCGACATCA AAAGATGGGA GAAAAAAATG GCGATGCAAA 840AACTTTCTGG ATGGAGCTAG AAGATGATGG AAAAGTGGAC TTCATTTTTG AACAAGTACA 900AAATGTGCTG CAGTCACTGA AACAAAAGAT CAAAGATGGG TCTGCCACCA ATAAAGAATA 960CATCCAAGCA ATGATTCTAG TGAATGAAGC AACTATAATT AACAGTTCAA CATCAATAAA 1020GGATCCTATG CCTGTGACTC AGAAGGAACA GGAAAACAAA TCCAATGCAT TTCCCTCTAC 1080ATCATGTGAA AACTCCTTTC CAGAAGACTG TACATTTCTA ACAACAGGAA ATAAGGAAAT 1140TCTCTCTCTT GAAGATAAAG TTGTAGACTT TAGAGAAAAA GACTCATCTT CGAATTTATC 1200TTACCAAAGT CATGACTGCT CTGGTGCTTG TCTGATGAAA ATGCCACTGA ACTTGAAGGG 1260AGAAAACCCT CTGCAGCTGC CAATCAAATG TCACTTCCAA AGACGACATG CAAAGACAAA 1320CTCTCATTCT TCAGCACTCC ACGTGAGTTA TAAAACCCCT TGTGGAAGGA GTCTACGAAA 1380CGTGGAGGAA GTTTTTCGTT ACCTGCTTGA GACAGAGTGT AACTTTTTAT TTACAGATAA 1440CTTTTCTTTC AATACCTATG TTCAGTTGGC TCGGAATTAC CCAAAGCAAA AAGAAGTTGT 1500TTCTGATGTG GATATTAGCA ATGGAGTGGA ATCAGTGCCC ATTTCTTTCT GTAATGAAAT 1560TGACAGTAGA AAGCTCCCAC AGTTTAAGTA CAGAAAGACT GTGTGGCCTC GAGCATATAA 1620TCTAACCAAC TTTTCCAGCA TGTTTACTGA TTCCTGTGAC TGCTCTGAGG GCTGCATAGA 1680CATAACAAAA TGTGCATGTC TTCAACTGAC AGCAAGGAAT GCCAAAACTT CCCCCTTGTC 1740AAGTGACAAA ATAACCACTG GATATAAATA TAAAAGACTA CAGAGACAGA TTCCTACTGG 1800CATTTATGAA TGCAGCCTTT TGTGCAAATG TAATCGACAA TTGTGTCAAA ACCGAGTTGT 1860CCAACATGGT CCTCAAGTGA GGTTACAGGT GTTCAAAACT GAGCAGAAGG GATGGGGTGT 1920ACGCTGTCTA GATGACATTG ACAGAGGGAC ATTTGTTTGC ATTTATTCAG GAAGATTACT 1980AAGCAGAGCT AACACTGAAA AATCTTATGG TATTGATGAA AACGGGAGAG ATGAGAATAC 2040TATGAAAAAT ATATTTTCAA AAAAGAGGAA ATTAGAAGTT GCATGTTCAG ATTGTGAAGT 2100TGAAGTTCTC CCATTAGGAT TGGAAACACA TCCTAGAACT GCTAAAACTG AGAAATGTCC 2160ACCAAAGTTC AGTAATAATC CCAAGGAGCT TACTATGGAA ACGAAATATG ATAATATTTC 2220AAGAATTCAG TATCATTCAG TTATTAGAGA TCCTGAATCC AAGACAGCCA TTTTTC 2276 ( 2 ) SEQ ID No.12

(ⅰ) sequence signature:

(A) length: 3114 base pairs

(B) type: nucleic acid

(C) chain: strand

( D ) ： ( ⅹⅰ ) SEQ ID NO:12：CAGGAGTCCG AACCCTTCAG TCATATAGAC CCAGAGGAGT CAGAGGAGAC CAGGCTCTTG 60AATATCTTAG GACTTATCTT CAAAGGCCCA GCAGCTTCCA CACAAGAAAA GAATCCCCGG 120GAGTCTACAG GAAACATGGT CACAGGACAG ACTGTCTGTA AAAATAAACC CAATATGTCG 180GATCCTGAGG AATCCAGGGG AAATGATGAA CTAGTGAAGC AGGAGATGCT GGTACAGTAT 240CTGCAGGATG CCTACAGCTT CTCCCGGAAG ATTACAGAGG CCATTGGCAT CATCAGCAAG 300ATGATGTATG AAAACACAAC TACAGTGGTG CAGGAGGTGA TTGAATNCTT TGTGATGGTC 360TTCCAATTTG GGGTACCCCA GGCCCTGTTT GGGGTGCGCC GTATGCTGCC TCTCATCTGG 420TCTAAGGAGC CTGGTGTCCG GGAAGCCGTG CTTAATGCCT ACCGCCAACT CTACCTCAAC 480CCCAAAGGGG ACTCTGCCAG AGCCAAGGCC CAGGCTTTGA TTCAGAATCT CTCTCTGCTG 540CTAGTGGATG CCTCGGTTGG GACCATTCAG TGTCTTGAGG AAATTCTCTG TGAGTTTGTG 600CAGAAGGATG AGTTGAAACC AGCAGTGACC CATCTGCTGT GGGAGCGGGC CACCGAGAAG 660GTCGCCTGCT GTCCTCTGGA GCGCTGTTCC TCTGTCATGC TTCTTGGCAT GATGGCACGA 720AGAAAGCCAG AAATTGTGGG AAGCAATTTA GACACACTGA TGAGCATAGG GCTGGATGAG 780AAGTTTCCAC AGGACTACAG GCTGGCCCAG CAGGTGTGCC ATGCCATTGC CAACATCTCG 840GACAGGAGAA AGCCTTCTCT GGGCAAACGT CACCCCCCCT TCCGGCTGCC TCAGGAACAC 900AGGTTGTTTG AGCGACTGCG GGAGACAGTC ACAAAAGGCT TTGTCCACCC AGACCCACTC 960TGGATCCCAT TCAAAGAGGT GGCAGTGACC CTCATTTACC AACTGGCAGA GGGCCCCGAA 1020GTGATCTGTG CCCAGATATT GCAGGGCTGT GCAAAACAGG CCCTGGAGAA GCTAGAAGAG 1080AAGAGAACCA GTCAGGAGGA CCCGAAGGAG TCCCCCGCAA TGCTCCCCAC TTTCCTGTTG 1140ATGAACCTGC TGTCCCTGGC TGGGGATGTG GCTCTGCAGC AGCTGGTCCA CTTGGAGCAG 1200GCAGTGAGTG GAGAGCTCTG CCGGCGCCGA GTTCTCCGGG AAGAACAGGA GCACAAGACC 1260AAAGATCCCA AGGAGAAGAA TACGAGCTCT GAGACCACCA TGGAGGAGGA GCTGGGGCTG 1320GTTGGGGCAA CAGCAGATGA CACAGAGGCA GAACTAATCC GTGGCATCTG CGAGATGGAA 1380CTGTTGGATG GCAAACAGAC ACTGGCTGCC TTTGTTCCAC TCTTGCTTAA AGTCTGTAAC 1440AACCCAGGCC TCTATAGCAA CCCAGACCTC TCTGCAGCTG CTTCACTTGC CCTTGGCAAG 1500TTCTGCATGA TCAGTGCCAC TTTCTGCGAC TCCCAGCTTC GTCTTCTGTT CACCATGCTG 1560GAAAAGTCTC CACTTCCCAT TGTCCGGTCT AACCTCATGG TTGCCACTGG GGATCTGGCC 1620ATCCGCTTTC CCAATCTGGT GGACCCCTGG ACTCCTCATC TGTATGCTCG CCTCCGGGAC 1680CCTGCTCAGC AAGTGCGGAA AACAGCGGGG CTGGTGATGA CCCACCTGAT CCTCAAGGAC 1740ATGGTGAAGG TGAAGGGGCA GGTCAGTGAG ATGGCGGTGC TGCTCATCGA CCCCGAGCCT 1800CAGATTGCTG CCCTGGCCAA GAACTTCTTC AATGAGCTCT CCCACAAGGG CAACGCAATC 1860TATAATCTCC TTCCAGATAT CATCAGCCGC CTGTCAGACC CCGAGCTGGG GGTGGAGGAA 1920GAGCCTTTCC ACACCATCAT GAAACAGCTC CTCTCCTACA TCACCAAGGA CAAGCAGACA 1980GAGAGCCTGG TGGAAAAGCT GTGTCAGCGG TTCCGCACAT CCCGAACTGA GCGGCAGCAG 2040CGAGACCTGG CCTACTGTGT GTCACAGCTG CCCCTCACAG AGCGAGGCCT CCGTAAGATG 2100CTTGACAATT TTGACTGTTT TGGAGACAAA CTGTCAGATG AGTCCATCTT CAGTGCTTTT 2160TTGTCAGTTG TGGGCAAGCT GCGACGTGGG GCCAAGCCTG AGGGCAAGGC TATAATAGAT 2220GAATTTGAGC AGAAGCTTCG GGCCTGTCAT ACCAGAGGTT TGGATGGAAT CAAGGAGCTT 2280GAGATTGGCC AAGCAGGTAG CCAGAGAGCG CCATCAGCCA AGAAACCATC CACTGGTTCT 2340AGGTACCAGC CTCTGGCTTC TACAGCCTCA GACAATGACT TTGTCACACC AGAGCCCCGC 2400CGTACTACCC GTCGGCATCC AAACACCCAG CAGCGAGCTT CCAAAAAGAA ACCCAAAGTT 2460GTCTTCTCAA GTGATGAGTC CAGTGAGGAA GATCTTTCAG CAGAGATGAC AGAAGACGAG 2520ACACCCAAGA AAACAACTCC CATTCTCAGA GCATCGGCTC GCAGGCACAG ATCCTAGGAA 2580GTCTGTTCCT GTCCTCCCTG TGCAGGGTAT CCTGTAGGGT GACCTGGAAT TCGAATTCTG 2640TTTCCCTTGT AAAATATTTG TCTGTCTCTT TTTTTTAAAA AAAAAAAAGG CCGGGCACTG 2700TGGCTCACGC CTGTAATCCC AGCACTTTGC GATACCAAGG CGGGTGGATA ACCTGAGGTA 2760GGGAGTTCGA GACCAGCCTG ACCAACATGG AGAAACCCCA TCTCTACTAA AAATAAAAAA 2820TTAGCCGGGC GTATTGGCGT GCGCCTGTAA TCCCAGCTAC TCAAGAGGCT GAGGCAGGAG 2880AATCGCCTGA ACCCAGAGGC GGAGGTTGTA GTGAGCCGAA ATCACACCAT TGCACTCCAG 2940CTTGGGCAAC AATAGCGAAC CTCCATCTCA AATTAAAAAA AAAATGCCTA CACGCTCTTT 3000AAAATGCAAG GCTTTCTCTT AAATTAGCCT AACTGAACTG CGTTGAGCTG CTTCAACTTT 3060GGAATATATG TTTGCCAATC TCCTTGTTTT CTAATGAATA AATGTTTTTA TATA 3114 ( 2 ) SEQ IDNo.13

(ⅰ) sequence signature:

(A) length: 1797 base pairs

(B) type: nucleic acid

(C) chain: strand

( D ) ： ( ⅹⅰ ) SEQ ID NO:13：CGGCACGAGA TCGACTGGTT GCAAGTAAAA CAGATGGAAA AATAGTACAG TATGAATGTG 60AGGGGGATAC TTGCCAGGAA GAGAAAATAG ATGCCTTACA GTTAGAGTAT TCATATTTAC 120TAACAAGCCA GCTGGAATCT CAGCGAATCT ACTGGGAAAA CAAGATAGTT CGGATAGAGA 180AGGACACAGC AGAGGAAATT AACAACATGA AGACCAAGTT TAAAGAAACA ATTGAGAAGT 240GTGATAATCT AGAGCACAAA CTAAATGATC TCCTAAAAGA AAAGCAGTCT GTGGAAAGAA 300AGTGCACTCA GCTAAACACA AAAGTGGCCA AACTCACCAA CGAGCTCAAA GAGGAGCAGG 360AAATGAACAA GTGTTTGCGA GCCAACCAAG TCCTCCTGCA GAACAAGCTA AAAGAGGAGG 420AGAGGGTGCT GAAGGAGACC TGTGACCAAA AAGATCTGCA GATCACCGAG ATCCAGGAGC 480AGCTGCGTGA CGTCATGTTC TACCTGGAGA CACAGCAGAA GATCAACCAT CTGCCTGCCG 540AGACCCGGCA GGAAATCCAG GAGGGACAGA TCAACATCGC CATGGCCTCG GCCTCGAGCC 600CTGCCTCTTC GGGGGGCAGT GGGAAGTTGC CCTCCAGGAA GGGCCGCAGC AAGAGGGGCA 660AGTGACCTTC AGAGCAACAG ACATCCCTGA GACTGTTCTC CCTGACACTG TGAGAGTGTG 720CTGGGACCTT CAGCTAAATG TGAGGGTGGG CCCTAATAAG TACAAGTGAG GATCAAGCCA 780CAGTTGTTTG GCTCTTTCAT TTGCTAGTGT GTGATGTANT GAATGTAAAG GGTGCTGACT 840GGAGAGCTGA TAGAAAGGCG CTGCGTTCGA AAAGGTCTTA ANAGTTCACT AACCTCACAT 900TCTAATGACC ATTTTGCCTT CCTGCTTGGT AGAAGCCCCA ACTCTGCTGT GCATTTTTCC 960ATTGTATTTA TGGAGTTGGC GTATTTGACA TTCAGTTCTG GGGTAGGTTT AAGATGTTAA 1020GTTATTTCTT GTAACCTCAA AGGTAAGGTT ATCTAGCACT AAAGCACCAA ACCTCTCTGA 1080GGGCATAACA GCTGCTTTAA AGAGAGGTTT CCATTGGCTA TTAAGGAGTT ATGAAAACTC 1140CCTAGCAATA GTGTCATATC ATTATCATCT CCCCCTTCCT CTGGGGAGTG GAAGAATTGC 1200TTGAATGTTA TCTGAAAAGA GGCCTGGTAG TAAACCAGGC CCTGGCTCTT TACCAGCAGT 1260CATCTCTTCT TGCTCTGGGG CCAGCCAGGA AAAACAAACA ACCCGGGGCA CATTGGGTAG 1320ACTCAGTGTA GGAAAAATGG TGGCAGCTCC ACTGTTTATT TTTGGTGACT TCGTACGTCA 1380TTATGAACCG CAATTAAGGA GGAGGCTTAA TGGCTGTTCC CAAACTCAAA TCTCAGAGTG 1440GGTATCCTAG CATCTAGCAA NACTGAGTGG GGAGATTTCT CATCCGTGTG AAAATGTAGA 1500GTGAGGCCTC TGACTAGCTN ATTGTGTATT TTGTTGGGTT TAGTATTTTC TAAATGTTTA 1560CAAAATATTG GGCTGCATGT TCAGGTTGCA GCTANAGGGA GCTTGGGCAN ATTTTCAATT 1620ACGCTTTCAA GATATAACCA AAAGCTGTTT CTAAATCCTA AAATTAGAAT TTCAACAGAN 1680CCCCCTTTAG AACAGTCATA TAACGCTTGT GTGGGCCAAC AGANGGGCTG TGTACTCTCT 1740CTGGAACCAT AAATGTCAAA TAATTTATAA CCTGCANTAA TTGAGCAACT TAAATAA 1797 ( 2 ) SEQ ID No.14

(ⅰ) sequence signature:

(A) length: 720 base pairs

(B) type: nucleic acid

(C) chain: strand

( D ) ： ( ⅹⅰ ) SEQ ID NO:14：TAATCACCAT CTGTTTTTGT GGGATGTGCT GCAGCATTTC CCAAAAAACT TNACGTGTAA 60TGTTGCAAAA TGAATGTACT CAGACATTNT TAATTTTTAC TTAGGGCAGA CCAACTCTTT 120GAGTCTCTCT TGGACTTATA TATACAGATA TCTTAAGAGT GGGAATGTAA AGCATAACCT 180AATTNTCTTT CCTATAGAGA TTCTATTTTA TTTAAAATNT ATTTNTACAC TAGTTAGAAT 240CCTGCTGTTT TGGCCAAGTA CTTGTCTTGC ATGTCTGACC TTGCAGAAGC TGGGGTGGAT 305CATAGCATAC TAATGAAGAG AATTAGAAGT AGTTTACAAA GCTCGCTCAC TCCTCATTTC 360TCTGTGATCC CTTCTATCCA GTGGCCCCAC CACCACCTGG GAAAACAGAT TTTTCAGTAC 420AGGTGGGATA AATGCTCTGA AAGGCTGTGC CCAGAGGAAT GAGCAAATAG GCAAGTGTTT 480CCAAACTACT TGGAGGTTTA CAAAAAATAT GTCCCAGAAA AAAAAAAAAT CTTACCAAGA 540TACGTAAAGA AAAAAAAATT TTTTTTTAAA CAGTCAAAGA GTCATGTTTG AATTTCACAA 600AATCACATCA GACAGAAGTT GTTTTCTTCA GGAGGGAAAT GAACCACTTA ATATACCCAT 660ACTACCTTGA ACAATGAAAT TGAATTAAAA TAGCCAAACT TTGAAAAAAA AAAAAAAAAA 720 ( 2 ) SEQ ID No.15

(ⅰ) sequence signature:

(A) length: 1996 base pairs

(B) type: nucleic acid

(C) chain: strand

( D ) ： ( ⅹⅰ ) SEQ ID NO:15：CAGAAGTGCA GCGGTGGCGG CGGCTGGTTG CGGGCCGGCG GCGGGCTGGC GGAGATGGAG 60GTAACTCAGG ATCTTGTTCA AGATGGGGTG GCTTCACCAG CTACCCCTGG GACCGGGAAA 120TCTAAGCTGG AAACATTGCC CAAAGAAGAC CTCATCAAGT TTGCCAAGAA ACAGATGATG 180CTAATACAGA AAGCTAAATC AAGGTGTACA GAATTGGAGA AAGAAATTGA AGAACTCAGA 240TCAAAACCTG TTACTGAAGG AACTGGTGAT ATTATTAAGG CATTAACTGA ACGTCTGGAT 300GCTCTTCTTC TGGAAAAAGC AGAGACTGAG CAACAGTGTC TTTCTCTGAA AAAGGAAAAT 360ATAAAAATGA AGCAAGAGGT TGAGGATTCT GTAACAAAGA TGGGAGATGC ACATAAGGAG 420TTGGAACAAT CACATATAAA CTATGTGAAA GAAATTGAAA ATTTGAAAAA TGAGTTGATG 480GCAGTACGTT CCAAATACAG TGAAGACAAA GCTAACTTAC AAAAGCAGCT GGAAGAACAA 540TGAATACGCA ATTAGAACTT TCAGAACAAC TTAAATTTCA GAACAACTCT GAAGATAATG 600TTAAAAAACT ACAAGAAGAG ATTGAGAAAA TTAGGCCAGG CTTTGAGGAG CAAATTTTAT 660ATCTGCAAAA GCAATTAGAC GCTACCACTG ATGAAAAGAA GGAAACAGTT ACTCAACTCC 720AAAATATCAT TGAGGCTAAT TCTCAGCATT ACCAAAAAAA TATTAATAGT TTGCAGGAAG 780AGCTTTTACA GTTGAAAGCT ATACACCAAG AAGAGGTGAA AGAGTTGATG TGCCAGATTG 840AAGCATCAGC TAAGGAACAT GAAGCAGAGA TAAATAAGTT GAACGAGCTA AAAGAGAACT 900TAGTAAAACA ATGTGAGGCA AGTGAAAAGA ACATCCAGAA GAAATATGAA TGTGAGTTAG 960AAAATTTAAG GAAAGCCACC TCAAATGCAA ACCAAGACAA TCAGATATGT TCTATTCTCT 1020TGCAAGAAAA TACATTTGTA GAACAAGTAG TAAATGAAAA AGTCAAACAC TTAGAAGATA 1080CCTTAAAAGA ACTTGAATCT CAACACAGTA TCTTAAAAGA TGAGGTAACT TATATGAATA 1140ATCTTAAGTT AAAACTTGAA ATGGATGCTC AACATATAAA GGATGAGTTT TTTCATGAAC 1200GGGAAGACTT AGAGTTTAAA ATTAATGAAT TATTACTAGC TAAAGAAGAA CAGGGCTGTG 1260TAATTGAAAA ATTAAAATCT GAGCTAGCAG GTTTAAATAA ACAGTTTTGC TATACTGTAG 1320AACAGCATAA CAGAGAAGTA CAGAGTCTTA AGGAACAACA TCAAAAAGAA ATATCAGAAC 1380TAAATGAGAC ATTTTTGTCA GATTCAGAAA AAGAAAAATT AACATTAATG TTTGAAATAC 1440AGGGTCTTAA GGAACAGTGT GAAAACCTAC AGCAAGAAAA GCAAGAAGCA ATTTTAAATT 1500ATGAGAGTTT ACGAGAGATT ATGGAAATTT TACAAACAGA ACTGGGGGAA TCTGCTGGAA 1560AAATAAGTCA AGAGTTCGAA TCAATGAAGC AACAGCAAGC ATCTGATGTT CATGAACTGC 1620AGCAGAAGCT CAGAACTGCT TTTACTGAAA AAGATGCCCT TCTCGAAACT GTGAATCGCC 1680TCCAGGGAGA AAATGAAAAG TTACTATCTC AACAAGAATT GGTACCAGAA CTTGAAAATA 1740CCATAAAGAA CCTTCAAGAA AAGAATGGAG TATACTTACT TAGTCTCAGT CAAAGAGATA 1800CCATGTTAAA AGAATTAGAA GGAAAGATAA ATTCTCTTAC TGAGGAAAAA GATGATTTTA 1860TAAATAAACT GAAAAATTCC CATGAAGAAA TGGATAATTT CCATAAGAAA TGTGAAAGGG 1920AAGAAAGATT GATTCTTGAA CTTGGGAAGA AAGTAGAGCA AACTATCCAG TACAACAGTG 1980AACTAGAACA AAAGGT 1996 ( 2 ) SEQ ID No.16

(ⅰ) sequence signature:

(A) length: 3642 base pairs

(B) type: nucleic acid

(C) chain: strand

( D ) ： ( ⅹⅰ ) SEQ ID NO:16：GTCCTGCTGA AGCTCACTCA GATTCCCTCA TTGATACCTT TCCTGAGTGT AGTACGGAAG 60GCTTCTCCAG TGACAGTGAT CTGGTATCTC TTACTGTTGA TGTGGATTCT CTTGCTGAGT 120TAGATGATGG AATGGCTTCC AATCAAAATT CTCCCATTAG AACTTTTGGT CTCAATCTTT 180CTTCGGATTC TTCAGCACTA GGGGCTGTTG CTTCTGACAG TGAACAGAGC AAAACAGAAG 240AAGAACGGGA AAGTCGTAGC CTCTTTCCTG GCAGTTTAAA GCCGAAGCTT GGCAAGAGAG 300ATTATTTGGA GAAAGCAGGA GAATTAATAA AGCTGGCTTT AAAAAAGGAA GAAGAAGACG 360ACTATGAAGC TGCTTCTGAT TTTTATAGGA AGGGAGTTGA TTTACTCCTA GAAGGTGTTC 420AAGGAGAGTC AAGCCCTACC CGTCGAGAAG CTGTGAAGAG AAGAACAGCC GAGTACCTCA 480TGCGGGCAGA AAGTATCTCT AGTCTTTATG GGAAACCTCA GCTTGATGAT GTATCTCAGC 540CTCCAGGATC ACTAAGTTCA AGGCCCCTTT GGAACCTAAG GAGCCCTGCC GAGGAGCTGA 600AGGCCTTCAG AGTCCTTGGG GTGATTGACA AGGTTTTACT TGTAATGGAC ACAAGGACAG 660AACACACTTT CATTTTAANA GGTCTAAGGA AAAGCAGTGA ATACAGCAGG AACAGAAAGA 720CCATCCNCCC CCGCTGTGTG CCCANCATGG TGTGTCTGCA TAAGTACATC ATCTCTGAAG 780AGTCANTATT TCTTGTGCTG CAGCATGCGG AANGTGGCAA ACTGTGGTCA TATATCAGTA 840AATTTCTAAA CAGAAGTCCT GAAGAAAGCT TTGACATCAA GGAAGTGAAA AAACCTACAC 900TTGCAAAAGT TCACCTGCAG CAGCCAACTT CTAGTCCTCA GGACAGCAGT AGCTTTGAAT 960CCAGAGGAAG TGATGGTGGA AGCATGCTTA AAGCTCTGCC TTTGAAGAGT AGTCTTACTC 1020CAAGTTCTCA AGATGACAGC AACCAGGAAG ATGATGGCCA AGATAGCTCT CCAAAGTGGC 1080CAGATTCTGG TTCAAGTTCA GAAGAAGAAT GTACTACTAG TTATTTAACA TTATGCAATG 1140AATATGGGCA AGAAAAGATT GAACCAGGGT CTTTGAATGA GGAGCCCTTC ATGAAGACTG 1200AAGGGAATGG TGTTGATACA AAAGCTATTA AAAGCTTCCC AGCACACCTT GCTGCTGACA 1260GTGACAGCCC CAGCACACAG CTGAGAGCTC ACGAGCTGAA GTTCTTCCCC AACGATGACC 1320CAGAAGCAGT TAGTTCTCCA AGAACATCAG ATTCCCTCAG TAGATCAAAA AATAGCCCCA 1380TGGAATTCTT TAGGATAGAC AGTAAGGATA GCGCAAGTGA ACTCCTGGGA CTTGACTTTG 1440GAGAAAAATT GTATAGTCTA AAATCAGAAC CTTTGAAACC ATTCTTTACT CTTCCAGATG 1500GAGACAGTGC TTCTAGGAGT TTTAATACTA GTGAAAGCAA GGTAGAGTTT AAAGCTCAGG 1560ACACCATTAG CAGGGGCTCA GATGACTCAG TGCCAGTTAT TTCATTTAAA GATGCTGCTT 1620TTGATGATGT CAGTGGTACT GATGAAGGAA GACCTGATCT TCTTGTAAAT TTACCTGGTG 1680AATTGGAGTC AACAAGAGAA GCTGCAGCAA TGGGACCTAC TAAGTTTACA CAAACTAATA 1740TAGGGATAAT AGAAAATAAA CTCTTGGAAG CCCCTGATGT TTTATGCCTC AGGCTTAGTA 1800CTGAACAATG CCAAGCACAT GAGGAGAAAG GCATAGAGGA ACTGAGTGAT CCCTCTGGGC 1860CCAAATCCTA TAGTATAACA GAGAAACACT ATGCACAGGA GGATCCCAGG ATGTTATTTG 1920TAGCANCTGT TGATCATAGT AGTTCAGGAG ATATGTCTTT GTTACCCAGC TCAGATCCTA 1980AGTTTCAAGG ACTTGGAGTG GTTGAGTCAN CAGTAACTGC AAACAACACA GAAGAAAGCT 2040TATTCCGTAT TTGTAGTCCA CTCTCAGGTG CTAATGAATA TATTGCAAGC ACAGACACTT 2100TAAAAACAGA AGAAGTATTG CTGTTTACAG ATCAGACTGA TGATTTGGCT AAAGAGGAAC 2160CAACTTCTTT ATTCCANAGA GACTCTGAGA CTAAGGGTGA AAGTGGTTTA GTGCTAGAAG 2220GAGACAAGGA AATACATCAG ATTTTTGAAG GACCTTGATA AAAAATTAGC ACTANCCTCC 2280AGGTTTTACA TCCCAGAGGG CTGCATTCAA AGNTGGGCAG CTGAAATGGT GGTAGCCCTT 2340NGATGCTTTA ACATAGAGAG GGAATTGTGT GCCGCGATTG AACCCAAACA ANATNTTATT 2400GAATGATAGA GGACACATTC AGNTAACGTA TTTTAGCAGG TGGAGTGAGG TTGAAGATTC 2460CTGTGACAGC GATGCCATAG AGAGAATGTA CTGTGCCCCA GAGGTTGGAG CAATCACTGA 2520AGAAACTGAA GCCTGTGATT GGTGGAGTTT GGGTGCTGTC CTCTTTGAAC TTNTCACTGG 2580CAAGACTCTG GTTGAATGCC ATCCAGCAGG AATAAATACT CACACTACTT TGAACATGCC 2640AGAATGTGTC TCTGAAGAGG CTCGCTCACT CATTCAACAG CTCTTGCAGT TCAATCCTCT 2700GGAACGACTT GGTGCTGGAG TTGCTGGTGT TGAAGATATC AAATCTCATC CATTTTTTAC 2760CCCTGTGGAT TGGGCAGAAC TGATGAGATG AACGTAATGC AGGGTTATCT TCACACATTC 2820TGATCTTCTC TGTGACAGGC ATCTCCAGCA CTGAGGCACC TCTGACTCAC AGTTACTTAT 2880GGAGCACCAA AGCATTTGGA TAAGGACCGT TATAGGAAAT GGGGGGGAAA TGGCTAAAAG 2940AGAACAATTT GTTTACAATT ACAAGATATT AGCTAATTGT GCCAGGGGCT GTTATATACA 3000TATATACACA ACCAAGGTGT GATCTGAATT TAATCCACAT TTGGTGTTGC AGATGAGTTG 3060TAAAGCCAAC TGAAAGAGTT CCTTCAAGAA GTTCCTCTGA TAGGAAGCTA GAAGTGTAGA 3120ATGAAGTTTT ACTTGACAGA AGGACCTTTA CATGGCAGCT AACAGTGCTT TTTGCTGACC 3180AGGATTGGTT TATATGATTA AATTAATATT TGCTTAATAA TACACTAAAA GTATATGAAC 3240AATGTCATCA ATGAAACTTA AAAGCGAGAA AAAAGAATAT ACACATAATT TCTGACGGAA 3300AACCTGTACC CTGATGCTGT ATAATGTATG TTGAATGTGG TCCCAGATTA TTTCTGTAAG 3360AAGACACTCC ATGTTGTCAG CTTTGTACTC TTTGTTGATA CTGCTTATTT AGAGAAGGGT 3420TCATATAAAC ACTCACTCTG TGTCTTCAAC AGCATCTTTC TTTCCCCATC TTTCTATTTT 3480CTGCACCCTC TGCTTGTTCC CTCATATTCT GTTCTTCCGA CTCCTGCTAA CACACATGCA 3540ACAAAAAAGG GAAGGGAGTG CTTATTTCCC TTTGTGTAAG GACTAAGAAA TCATGATATC 3600AAATAAACAT GGTGAAACAT TNANAAAAAA AAAAAAAAAA AA 3642 ( 2 ) SEQ ID No.17

(ⅰ) sequence signature:

(A) length: 1397 base pairs

(B) type: nucleic acid

(C) chain: strand

( D ) ： ( ⅹⅰ ) SEQ ID NO:17：GTTCAACTCA ATAGAAGATG ACGTTTGCCA GCTAGTGTAT GTGGAAAGAG CTGAAGTGCT 60CAAATCTGAA GATGGCGCCA GCCTCCCAGT GATGGACCTG ACTGAACTCC CCAAGTGCAC 120GGTGTGTCTG GAGCGCATGG ACGAGTCTGT GAATGGCATC CTCACAACGT TATGTAACCA 180CATCTTCCAC AGCCAGTGTC TACAGCGCTG GGACGATACC ACGTGTCCTG TTTGCCGGTA 240CTGTCAAACG CCCGAGCCAG TAGAAGAAAA TAAGTGTTTT GAGTGTGGTG TTCAGGAAAA 300TCTTTGGATT TGTTTAATAT GCGGCCACAT AGGATGTGGA CGGTATGTCA GTCGACATGC 360TTATAAGCAC TTTGAGGAAA CGCAGCACAC GTATGCCATG CAGCTTACCA ACCATCGAGT 420CTGGGACTAT GCTGGAGATA ACTATGTTCA TCGACTGGTT GCAAGTAAAA CAGATGGAAA 480AATAGTACAG TATGAATGTG AGGGGGATAC TTGCCAGGAA GAGAAAATAG ATGCCTTACA 540GTTAGAGTAT TCATATTTAC TAACAAGCCA GCTGGAATCT CAGCGAATCT ACTGGGAAAA 600CAAGATAGTT CGGATAGAGA AGGACACAGC AGAGGAAATT AACAACATGA AGACCAAGTT 660TAAAGAAACA ATTGAGAAGT GTGATAATCT AGAGCACAAA CTAAATGATC TCCTAAAAGA 720AAAGCAGTCT GTGGAAAGAA AGTGCACTCA GCTAAACACA AAAGTGGCCA AACTCACCAA 780CGAGCTCAAA GAGGAGCAGG AAATGAACAA GTGTTTGCGA GCCAACCAAG TCCTCCTGCA 840GAACAAGCTA AAAGAGGAGG AGAGGGTGCT GAAGGAGACC TGTGACCAAA AAGATCTGCA 900GATCACCGAG ATCCAGGAGC AGCTGCGTGA CGTCATGTTC TACCTGGAGA CACAGCAGAA 960AGATCAACCA TCTGCCTGCC GAGACCCGGC AGGAAATCCA GGAGGGACAG ATCAACATCG 1020CCATGGCCTC GGCCTCGAGC CCTGCCTCTT CGGGGGGCAG TGGGAAGTTG CCCTCCAGGA 1080AGGGCCGCAG CAAGAGGGGC AAGTGACCTT CAGAGCAACA GACATCCCTG AGACTGTTCT 1140CCCTGACACT GTGAGAGTGT GCTGGGACCT TCAGCTAAAT GTGAGGGTGG GCCCTAATAA 1200GTACAAGTGA GGATCAAGCC ACAGTTGTTT GGCTCTTTCA TTTGCTAGTG TGTGATGTAG 1260TGAATGTAAA GGGTGCTGAC TGGAGAGCTG ATAGAAAGGC GCTGCGTTCG AAAAGGTCTT 1320AAGAGTTCAC TAACCTCACA TTCTAATGAC CANTTTGCCT TCCTGCTTGG TAGAAGCCCC 1380ACACTCTGCT GTGCATT 1397 ( 2 ) SEQ ID No.18

(ⅰ) sequence signature:

(A) length: 800 base pairs

(B) type: nucleic acid

(C) chain: strand

( D ) ： ( ⅹⅰ ) SEQ ID NO:18：CGGTAATTGA GCANACTTAA AATAAGACCT GTGTTGGAAT TTAGTTTCCT CTGAAGAGGT 60AGAGGGATAG GTTAGTAAGA TGTATTGTTA AACAACAGGT TTTAGTTTTT GCTTTTATAA 120TTAGCCACAG GTTTTCAAAT GATCACATTT CAGAATAGGT TTTTAGCCTG TAATTAGGCC 180TCATCCCCTT TGACCTAAAT GTCTTACATG TTACTTGTTA GCACATCAAC TGTATCACTA 240ATCACCATCT GNTTTTGTGG GATGTGCTGC AGCATTTCCC AAAAAACTTT ACGTGTAATG 300TTGCAAAATG AATGTACTCA GACATTCTTA ATTTTTACTT AGGGCAGACC AACTCTTTGA 360GTCTCTCTTG GACTTATATA TACAGATATC TTAAGAGTGG GAATGTAAAG CATAACCTAA 420TTCTCTTTCC TATAGAGATT CTATTTTATT TAAAATCTAT TTTTACACTA GTTAGAATCC 480TGCTGTTTTG GCCAAGTACT TGTCTTGCAT GTCTGACCTT GCAGAAGCTG GGGTGGATCA 540TAGCATACTA ATGAAGAGAA TTAGAAGTAG TTTACAAAGC TCGCTCACTC CTCATTTCTC 600TGTGATCCCT TCTATCCAGT GGCCCCACCA CCACCTGGGA AAACAGATTT TTCAGTACAG 660GTGGGATAAA TGCTCTGAAA GGCTGTGCCC AGAGGAATGA GCAAATAGGC AAGTGTTTCC 720AAACTACTTG GAGGTTTACA AAAAATATGT CCCAGAAAAA AAAAAAATCT TACCAAGATA 780CGTAAAAAAA AAAAAAAAAA 800 ( 2 ) SEQ ID No.19

(ⅰ) sequence signature:

(A) length: 1810 base pairs

(B) type: nucleic acid

(C) chain: strand

( D ) ： ( ⅹⅰ ) SEQ ID NO:19：GCAGCTCCCA GGTGCGTGTT AAAAGCTGGA GGGGGGATAT GTGATCCCAG GACCAAAAGC 60GCGGGGCCAG ACTCATCGGT TCATTCAACA ACCAGTATTT AGTGCCTGCT GTGTTCTGCA 120GGCCCTGCCA TAGGCGCTTG ATACAGCGGT GCATAGCGTA TGAAAAAGAT CTGTCCTGGC 180TGAGCATCCG TAATATAAAA ATCTGAAATC TGAAATGCTC CAAAATCCTA AACTTTTTGA 240GTGCTGACAT TATGCCACAA ATGGAAAATT TCATACCTGA CCTTATGTGG GTTGCANTCA 300AAACACAGGT GCACAACACC CAGTTCATGC AACATCCCCA ATGGGAAAAA AGACCCCCCC 360AGCTCTCTTC TGCTGCAGTT TTTCTGCTCA CACCTGGATT TCCCCATGCA TTCCCACAAA 420AAGTAATTAA ATGGCATGCG TGCAGGCTGG ACACGCCAAC AACAGGTTTC CCACAATGCC 480CCACATGGGG CCAAGACCTG TGTGCATTAC TCATTGCATT TTTTTGCTTA TTCTCTGCTG 540TGTGGTATAA ATATATTGTT GAAAATGTCA AAAAGACCTA AAGATACCCC TGTGAATATC 600AGTGATAAGA AAAAGAGGAA GCATTTATGT TTATCTATAG CACAGAAAGT CAAGTTGTTG 660GAGAAACTGG ACAGTGGTGT AAGTGTGAAA CATCTTACAG AAGAGTATGG TGTTGGAATG 720ACCACCATAT ATGACCTGAA GAAACAGAAG GATAAACTGT TGAAGTTTTA TGCTGAAAGT 780GATGAGCAGA TATTAATGAA AAATAGAAAA ACACTTCATA AAGCTAAAAA TGAAGATCTT 840GATCGTGTAT TGAAAGAGTG GATCCGTCAG CGTCGCAGTG AACACATGCC ACTTAATGGT 900ATGCTGATCA TGAAACAAGC AAAGATATAT CACAATGAAC TAAAAATTGA GGGGAACTGT 960GAATATTCAA CAGGCTGGTT GCAGAAATTT AAGAAAAGAC ATGGCATTAA ATTTTTAAAG 1020ACTTGTGGCA ATAAAGCATC TGCTGGTCAT GAAGCAACAG AGAAGTTTAC TGGCAATTTC 1080AGTAATGATG ATGAACAAGA TGGTAACTTT GAAGGATTCA NTATGTCAAG TGAGAAAAAA 1140ATAATGTCTG ACCTCCTTAC ATATACAAAA AATATACATC CAGAGACTGT CAGTAAGCTG 1200GAAGAAGAGG ATATCTTTNA TGTTTTTAAC AGTAATAATG AGGCTCCAGT TGTTCATTCA 1260TTGTCCAATG GTGAAGTAAC AAAAATGGTT CTGAATCAAG ATGATCATGA TGATAATGAT 1320AATGAAGATG ATGTTAACAC TGCAGAAAAA GTGCCTATAG ACGACATGGT AAAAATGTGT 1380GATGGGCTTA TTAAAGGACT AGAGCAGCAT GCATTCATAA CAGAGCAAGA AATCATGTCA 1440GTTTATAAAA TCAAAGAGAG ACTTCTAAGA CAAAAAGCAT CATTAATGAG GCAGATGACT 1500CTGAAAGAAA CATTTAAAAA AGCCATCCAG AGGAATGCTT CTTCCTCTCT ACAGGACCCA 1560CTTCTTGGTC CCTCAACTGC TTCTGATGCT TCTTCTCACC TAAAAATAAA ATAAAATACA 1620GTGTACAGTA ACCTTTTAGT CAAAACAGCA TCATACTTGG AAACTGAAAG CCTACTGTTA 1680TTTGTTATTG TTGCTTAACA GCTGATACAG GTATTCTGGT GACACTACTG TGCTGGCTTA 1740CTTAACCTGA ATACACTATT TTTTTCGTTG TAAAAAAAAA AAAAAAANAA NAAAAAAAAA 1800AAAAAANANA 1810 ( 2 ) SEQ ID No.20

(ⅰ) sequence signature:

(A) length: 70 amino acid

(B) type: amino acid

(C) chain:

(D) topological classification: the sequence description of linear (ⅹ ⅰ) SEQ ID NO:20: Ala Arg Glu Gly Gly Lys Met Val Leu Glu Ser Thr Met Val Cys Val1 5 10 15Asp Asn Ser Glu Tyr Met Arg Asn Gly Asp Phe Leu Pro Thr Arg Leu

20 25 30Gln?Ala?Gln?Gln?Asp?Ala?Val?Asn?Ile?Xaa?Cys?His?Ser?Lys?Thr?Arg

35 40 45Ser?Asn?Pro?Glu?Asn?Asn?Val?Gly?Leu?Ile?Thr?Leu?Ala?Asn?Asp?Cys

The information of 50 55 60Glu Val Leu Thr Thr Leu, 65 70 (2) SEQ ID No.21

(ⅰ) sequence signature:

(A) length: 100 amino acid

(B) type: amino acid

(C) chain:

(D) topological classification: the sequence description of linear (ⅹ ⅰ) SEQ ID NO:21: Ala Arg Glu Ser Thr Met Val Cys Val Asp Asn Ser Glu Tyr Met Arg1 5 10 15Asn Gly Asp Phe Leu Pro Thr Arg Leu Gln Ala Gln Gln Asp Ala Val

20 25 30Asn?Ile?Val?Cys?His?Ser?Lys?Thr?Arg?Ser?Asn?Pro?Glu?Asn?Asn?Val

35 40 45Gly?Leu?Ile?Thr?Leu?Ala?Asn?Asp?Cys?Glu?Val?Leu?Thr?Thr?Leu?Thr

50 55 60Pro?Asp?Thr?Gly?Arg?Ile?Leu?Ser?Lys?Leu?His?Thr?Val?Gln?Pro?Lys65 70 75 80Gly?Lys?Ile?Thr?Phe?Cys?Thr?Gly?Ile?Arg?Val?Ala?His?Leu?Ala?Leu

85 90 95Lys?His?Arg?Gln

The information of 100 (2) SEQ ID No.22

(ⅰ) sequence signature:

(A) length: 214 base pairs

(B) type: nucleic acid

(C) chain: strand

(D) topological classification: linearity, the sequence description of (ⅹ ⅰ) SEQ ID NO:22: CGGCACGAGA AGGTGGCAAG ATGGTGTTGG AAAGCACTAT GGTGTGTGTG GACAACAGTG 60AGTATATGCG GAATGGAGAC TTCTTACCCA CCAGGCTGCA GGCCCAGCAG GATGCTGTCA 120ACATANTTTG TCATTCAAAG ACCCGCAGCA ACCCTGAGAA CAACGTGGGC CTTATCACAC 180TGGCTAATGA CTGTGAAGTG CTGACCACAC TCAC 214, (2) information of SEQ ID No.23

(ⅰ) sequence signature:

(A) length: 375 base pairs

(B) type: nucleic acid

(C) chain: strand

(D) topological classification: the sequence description of linear (ⅹ ⅰ) SEQ ID NO:23: the information of TATGGACACA TTTGAGCCAG CCAAGGAGGA GGATGATTAC GACGTGATGC AGGACCCCGA 60GTTCCTTCAG AGTGTCCTAG AGAACCTCCC AGGTGTGGAT CCCAACAATG AAGCCATTCG 120AAATGNTATG GGCTCCCTGG CCTCCCAGGC CACCAAGGAC GGCAAGAAGG ACAAGAAGGA 180GGAAGACAAG AAGTGAGACT GGAGGGAAAG GGTAGCTGAG TCTGCTTAGG GGACTGCATG 240GGAAGCACGG AATATAGGGT TAGATGTGTG TTATCTGTAA CCATTACAGC CTAAATAAAG 300CTTGGCAACT TTTTAAAAAA AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA 360AAAAAAAAAC TCGAG 375 (2) SEQ ID No.24

(ⅰ) sequence signature:

(A) length: 304 base pairs

(B) type: nucleic acid

(C) chain: strand

(D) topological classification: the sequence description of linear (ⅹ ⅰ) SEQ ID NO:24: the information of CGGCACGAGA AAGCACTATG GTGTGTGTGG ACAACAGTGA GTATATGCGG AATGGAGACT 60TCTTACCCAC CAGGCTGCAG GCCCAGCAGG ATGCTGTCAA CATAGTTTGT CATTCAAAGA 120CCCGCAGCAA CCCTGAGAAC AACGTGGGCC TTATCACACT GGCTAATGAC TGTGAAGTGC 180TGACCACACT CACCCCAGAC ACTGGCCGTA TCCTGTCCAA GCTACATACT GTCCAACCCA 240AGGGCAAGAT CACCTTCTGC ACGGGCATCC GCGTTGCCCA TCTGGCTCTG AAGCACCGAC 300AAGG 304 (2) SEQ ID No.25

(ⅰ) sequence signature:

(A) length: 22 amino acid

(B) type: amino acid

(C) chain:

(D) topological classification: the sequence description of linear (ⅹ ⅰ) SEQ ID NO:25: Val Arg Gly Gly Gly Gly Gly Gly Pro Gly Gly Gly Gly Val Gly Gly1 5 10 15Arg Cys Gly Gly Gly Gly

The information of 20 (2) SEQ ID No.26

(ⅰ) sequence signature:

(A) length: 78 amino acid

(B) type: amino acid

(C) chain:

(D) topological classification: the sequence description of linear (ⅹ ⅰ) SEQ ID NO:26: Ala Arg Ala Ala Arg Ala Lys Ala Gln Ala Leu Ile Gln Asn Leu Ser1 5 10 15Leu Leu Leu Val Asp Ala Ser Val Gly Thr Ile Gln Cys Leu Glu Glu

20 25 30Ile?Leu?Cys?Glu?Phe?Val?Gln?Lys?Asp?Glu?Leu?Lys?Pro?Ala?Val?Thr

35 40 45Xaa?Leu?Leu?Trp?Glu?Arg?Ala?Thr?Glu?Lys?Val?Ala?Cys?Cys?Pro?Leu

The information of 50 55 60Glu Arg Cys Ser Ser Val Met Leu Leu Gly Met Met Ala Arg65,70 75 (2) SEQ ID No.27

(ⅰ) sequence signature:

(A) length: 384 amino acid

(B) type: amino acid

(C) chain:

(D) topological classification: the sequence description of linear (ⅹ ⅰ) SEQ ID NO:27: Lys Met Val Leu Glu Ser Thr Met Val Cys Val Asp Asn Ser Glu Tyr1 5 10 15Met Arg Asn Gly Asp Phe Leu Pro Thr Arg Leu Gln Ala Gln Gln Asp

20 25 30Ala?Val?Asn?Ile?Val?Cys?His?Ser?Lys?Thr?Arg?Ser?Asn?Pro?Glu?Asn

35 40 45Asn?Val?Gly?Leu?Ile?Thr?Leu?Ala?Asn?Asp?Cys?Glu?Val?Leu?Thr?Thr

50 55 60Leu?Thr?Pro?Asp?Thr?Gly?Arg?Ile?Leu?Ser?Lys?Leu?His?Thr?Val?Gln65 70 75 80Pro?Lys?Gly?Lys?Ile?Thr?Phe?Cys?Thr?Gly?Ile?Arg?Val?Ala?His?Leu

85 90 95Ala?Leu?Lys?His?Arg?Gln?Gly?Lys?Asn?His?Lys?Met?Arg?Ile?Ile?Ala

100 105 110Phe?Val?Gly?Ser?Pro?Val?Glu?Asp?Asn?Glu?Lys?Asp?Leu?Val?Lys?Leu

115 120 125Ala?Lys?Arg?Leu?Lys?Lys?Glu?Lys?Val?Asn?Val?Asp?Ile?Ile?Asn?Phe

130 135 140Gly?Glu?Glu?Glu?Val?Asn?Thr?Glu?Lys?Leu?Thr?Ala?Phe?Val?Asn?Thr145 150 155 160Leu?Asn?Gly?Lys?Asp?Gly?Thr?Gly?Ser?His?Leu?Val?Thr?Val?Pro?Pro

165 170 175Gly?Pro?Ser?Leu?Ala?Asp?Ala?Leu?Ile?Ser?Ser?Pro?Ile?Leu?Ala?Gly

180 185 190Glu?Gly?Gly?Ala?Met?Leu?Gly?Leu?Gly?Ala?Ser?Asp?Phe?Glu?Phe?Gly

195 200 205Val?Asp?Pro?Ser?Ala?Asp?Pro?Glu?Leu?Ala?Leu?Ala?Leu?Arg?Val?Ser

210 215 220Met?Glu?Glu?Gln?Arg?Gln?Arg?Gln?Glu?Glu?Glu?Ala?Arg?Arg?Ala?Ala225 230 235 240Ala?Ala?Ser?Ala?Ala?Glu?Ala?Gly?Ile?Ala?Thr?Thr?Gly?Thr?Glu?Asp

245 250 255Ser?Asp?Asp?Ala?Leu?Leu?Lys?Met?Thr?Ile?Ser?Gln?Gln?Glu?Phe?Gly

260 265 270Arg?Thr?Gly?Leu?Pro?Asp?Leu?Ser?Ser?Met?Thr?Glu?Glu?Glu?Gln?Ile

275 280 285Ala?Tyr?Ala?Met?Gln?Met?Ser?Leu?Gln?Gly?Ala?Glu?Phe?Gly?Gln?Ala

290 295 300Glu?Ser?Ala?Asp?Ile?Asp?Ala?Ser?Ser?Ala?Met?Asp?Thr?Ser?Glu?Pro305 310 315 320Ala?Lys?Glu?Glu?Asp?Asp?Tyr?Asp?Val?Met?Gln?Asp?Pro?Glu?Phe?Leu

325 330 335Gln?Ser?Val?Leu?Glu?Asn?Leu?Pro?Gly?Val?Asp?Pro?Asn?Asn?Glu?Ala

340 345 350Ile?Arg?Asn?Ala?Met?Gly?Ser?Leu?Pro?Pro?Arg?Pro?Pro?Arg?Thr?Ala

355 360 365Arg?Arg?Thr?Arg?Arg?Arg?Lys?Thr?Arg?Ser?Glu?Thr?Gly?Gly?Lys?Gly

The information of 370 375 380 (2) SEQ ID No.28

(ⅰ) sequence signature:

(A) length: 68 amino acid

(B) type: amino acid

(C) chain:

(D) topological classification: linearity

The sequence description of (ⅹ ⅰ) SEQ ID NO:28: Ala Arg Asp Ala Tyr Ser Phe Ser Arg Lys Ile Thr Glu Ala Ile Gly1 5 10 15Ile Ile Ser Lys Met Met Tyr Glu Asn Thr Thr Thr Val Val Gln Glu

20 25 30Val?Ile?Glu?Phe?Phe?Val?Met?Val?Phe?Gln?Phe?Gly?Val?Pro?Gln?Ala

35 40 45Leu?Phe?Gly?Val?Arg?Arg?Met?Leu?Pro?Leu?Ile?Trp?Ser?Lys?Glu?Pro

The information of 50 55 60Gly Val Arg Glu65 (2) SEQ ID No.29

(ⅰ) sequence signature:

(A) length: 97 amino acid

(B) type: amino acid

(C) chain:

(D) topological classification: the sequence description of linear (ⅹ ⅰ) SEQ ID NO:29: Ala Arg Ala Gln Ala Leu Phe Gly Val Arg Arg Met Leu Pro Leu Ile1 5 10 15Trp Ser Lys Glu Pro Gly Val Arg Glu Ala Val Leu Asn Ala Tyr Arg

20 25 30Gln?Leu?Tyr?Leu?Ash?Pro?Lys?Gly?Asp?Ser?Ala?Arg?Ala?Lys?Ala?Gln

35 40 45Ala?Leu?Ile?Gln?Asn?Leu?Ser?Leu?Leu?Leu?Val?Asp?Ala?Ser?Val?Gly

50 55 60Thr?Ile?Gln?Cys?Leu?Glu?Glu?Ile?Leu?Cys?Glu?Phe?Val?Gln?Lys?Asp65 70 75 80Glu?Leu?Lys?Pro?Ala?Val?Thr?Gln?Leu?Leu?Trp?Glu?Pro?Ala?Thr?Glu

The information of 85 90 95Lys (2) SEQ ID No.30

(ⅰ) sequence signature:

(A) length: 116 amino acid

(B) type: amino acid

(C) chain:

(D) topological classification: the sequence description of linear (ⅹ ⅰ) SEQ ID NO:30: Ala Arg Ala Thr Thr Ala Phe Gly Cys Arg Ile Trp Asn Pro Cys Ala1 5 10 15Ala Leu Thr Met Lys Gln Ser Ser Asn Val Pro Ala Phe Leu Ser Lys

20 25 30Leu?Trp?Thr?Leu?Val?Glu?Glu?Thr?His?Thr?Asn?Glu?Phe?Ile?Thr?Trp

35 40 45Ser?Gln?Asn?Gly?Gln?Ser?Phe?Leu?Val?Leu?Asp?Glu?Gln?Arg?Phe?Ala

50 55 60Lys?Glu?Ile?Leu?Pro?Lys?Tyr?Phe?Lys?His?Asn?Asn?Met?Ala?Ser?Phe65 70 75 80Val?Arg?Gln?Leu?Asn?Met?Tyr?Gly?Phe?Arg?Lys?Val?Ile?His?Ile?Asp

85 90 95Ser?Gly?Ile?Val?Lys?Gln?Glu?Arg?Asp?Gly?Pro?Val?Glu?Phe?Gln?His

100 105 110Pro?Tyr?Phe?Gln

The information of 115 (2) SEQ ID No.31

(ⅰ) sequence signature:

(A) length: 124 amino acid

(B) type: amino acid

(C) chain:

(D) topological classification: the sequence description of linear (ⅹ ⅰ) SEQ ID NO:31: Ala Arg Gly Ala Thr Cys Glu Arg Cys Lys Gly Gly Phe Ala Pro Ala1 5 10 15Glu Lys Ile Val Asn Ser Asn Gly Glu Leu Tyr His Glu Gln Cys Phe

20 25 30Val?Cys?Ala?Gln?Cys?Phe?Gln?Gln?Phe?Pro?Glu?Gly?Leu?Phe?Tyr?Glu

35 40 45Phe?Glu?Gly?Arg?Lys?Tyr?Cys?Glu?His?Asp?Phe?Gln?Met?Leu?Phe?Ala

50 55 60Pro?Cys?Cys?His?Gln?Cys?Gty?Glu?Phe?Ile?Ile?Gly?Arg?Val?Ile?Lys65 70 75 80Ala?Met?Asn?Asn?Ser?Trp?His?Pro?Glu?Cys?Phe?Arg?Cys?Asp?Leu?Cys

85 90 95Gln?Glu?Val?Leu?Ala?Asp?Ile?Gly?Phe?Val?Lys?Asn?Ala?Gly?Arg?His

100 105 110Leu?Cys?Arg?Pro?Cys?His?Asn?Arg?Glu?Lys?Ala?Arg

The information of 115 120 (2) SEQ ID No.32

(ⅰ) sequence signature:

(A) length: 768 base pairs

(B) type: nucleic acid

(C) chain: strand

( D ) ： ( ⅹⅰ ) SEQ ID NO:32：TACGAGGAGG AGGAGGAGGA GGCCCCGGAG GAGGAGGCGT TGGAGGTCGA TGCGGAGGCG 60GAGGATGAGG AGGCCGAGGC GCCGGAGGAG GCCGAGGCGC CGGAGCAGGA GGAGGCCGGC 120CGGAGGCGGC ATGAGACGAG CGTGGCGGCC GCGGCTGCTC GGGGCCGCGC TGGTTGCCCA 180TTGACAGCGG CGTCTGCAGC TCGCTTCAAG ATGGCCGCTT GGCTCGCATT CATTTTCTGC 240TGAACGACTT TTAACTTTCA TTGTCTTTTC CGCCCGCTTC GATCGCCTCG CGCCGGCTGC 300TCTTTCCGGG ATTTTTTATC AAGCAGAAAT GCATCGAACA ACGAGAATCA AGATCACTGA 360GCTAAATCCC CACCTGATGT GTGTGCTTTG TGGAGGGTAC TTCATTGATG CCACAACCAT 420AATAGAATGT CTACATTCCT TCTGTAAAAC GTGTATTGTT CGTTACCTGG AGACCAGCAA 480GTATTGTCCT ATTTGTGATG TCCAAGTTCA CAAGACCAGA CCACTACTGA ATATAAGGTC 540AGATAAAACT CTCCAAGATA TTGTATACAA ATTAGTTCCA GGGCTTTTCA AAAATGAAAT 600GAAGAGAAGA AGGGATTTTT ATGCAGCTCA TCCTTCTGCT GATGCTGCCA ATGGCTCTAA 660TGAAGATNGA GGAGAGGTTG CAGATGAAGA TAAGAGAATT ATAACTGATG ATGAGATAAT 720AAGCTTATCC ATTGAATTCT TTGACCAGAA CAGATTGGAT CGGAAAGT 768 ( 2 ) SEQ ID No.33

(ⅰ) sequence signature:

(A) length: 642 base pairs

(B) type: nucleic acid

(C) chain: strand

( D ) ： ( ⅹⅰ ) SEQ ID NO:33：TTTAAATAAA CCAGCAGGTT GCTAAAAGAA GGCATTTTAT CTAAAGTTAT TTTAATAGGT 60GGTATAGCAG TAATTTTAAA TTTAAGAGTT GCTTTTACAG TTAACAATGG AATATGCCTT 120CTCTGCTATG TCTGAAAATA GAAGNTATTT ATTATGAGCT TNTACAGGTA TTTTTAAATA 180GAGCAAGCAT GTTGAATTTA AAATATGAAT AACCCCACCC AACAATTTTC AGTTTATTTT 240TTGCTTTGGT CGAACTTGGT GTGTGTTCAT CACCCATCAG TTATTTGTGA GGGTGTTTAT 300TCTATATGAA TATTGTTTCA TGTTTGTATG GGAAAATTGT AGCTAAACAT TTCATTGTCC 360CCAGTCTGCA AAAGAAGCAC AATTCTATTG CTTTGTCTTG CTTATAGTCA TTAAATCATT 420ACTTTTACAT ATATTGCTGT TACTTCTGCT TTCTTTAAAA ATATAGTAAA GGATGTTTTA 480TGAAGTCACA AGATACATAT ATTTTTATTT TGACCTAAAT TTGTACAGTC CCATTGTAAG 540TGTTGTTTCT AATTATAGAT GTAAAATGAA ATTTCATTTG TAATTGGAAA AAATCCAATA 600AAAAGGATAT TCATTTAAAA AAAAAAAAAA AAAAAAAAAA AA 642 ( 2 ) SEQ ID No.34

(ⅰ) sequence signature:

(A) length: 236 base pairs

(B) type: nucleic acid

(C) chain: strand

(D) topological classification: the sequence description of linear (ⅹ ⅰ) SEQ ID NO:34: the information of CGGCACGAGC TGCCAGAGCC AAGGCCCAGG CTTTGATTCA GAATCTCTCT CTGCTGCTAG 60TGGATGCCTC GGTTGGGACC ATTCAGTGTC TTGAGGAAAT TCTCTGTGAG TTTGTGCAGA 120AGGATGAGTT GAAACCAGCA GTGACCCANC TGCTGTGGGA GCGGGCCACC GAGAAAGTCG 180CCTGCTGTCC TCTGGAACGC TGTTCCTCTG TCATGCTTCT TGGCATGATG GCACGA 236 (2) SEQ ID No.35

(ⅰ) sequence signature:

(A) length: 333 base pairs

(B) type: nucleic acid

(C) chain: strand

(D) topological classification: the sequence description of linear (ⅹ ⅰ) SEQ ID NO:35: the information of CCGGGCGTAT TGGCGTGCGC CTGTAATCCC AGCTAACTCA AGAGGCTGAG GCAGGAGAAT 60CGCCTGAACC CAGAGGCGGA GGTTGTAGTG AGCCGAAATC ACACCATTGC ACTCCAGCTT 120GGGCAACAAT AGCGAACCTC CATCTCAAAT TAAAAAAAAA AATGCCTACA CGCTCTTTAA 180AATGCAAGGC TTTCTCTTAA ATTAGCCTAA CTGAACTGCG TTGAGCTGCT TCAACTTTGG 240AATATATGTT TGCCAATCTC CTTGTTTTCT AATGAATAAA TGTTTTTATA TACTTTTAGA 300AAAAAAAAAA AAAAAAAAAA AAAAAAACTC GAG 333 (2) SEQ ID No.36

(ⅰ) sequence signature:

(A) length: 1272 base pairs

(B) type: nucleic acid

(C) chain: strand

( D ) ： ( ⅹⅰ ) SEQ ID NO:36：GCAAGATGGT GTTGGAAAGC ACTATGGTGT GTGTGGACAA CAGTGAGTAT ATGCGGAATG 60GAGACTTCTT ACCCACCAGG CTGCAGGCCC AGCAGGATGC TGTCAACATA GTTTGTCATT 120CAAAGACCCG CAGCAACCCT GAGAACAACG TGGGCCTTAT CACACTGGCT AATGACTGTG 180AAGTGCTGAC CACACTCACC CCAGACACTG GCCGTATCCT GTCCAAGCTA CATACTGTCC 240AACCCAAGGG CAAGATCACC TTCTGCACGG GCATCCGCGT GGCCCATCTG GCTCTGAAGC 300ACCGACAAGG CAAGAATCAC AAGATGCGCA TCATTGCCTT TGTGGGAAGC CCAGTGGAGG 360ACAATGAGAA GGATCTGGTG AAACTGGCTA AACGCCTCAA GAAGGAGAAA GTAAATGTTG 420ACATTATCAA TTTTGGGGAA GAGGAGGTGA ACACAGAAAA GCTGACAGCC TTTGTAAACA 480CGTTGAATGG CAAAGATGGA ACCGGTTCTC ATCTGGTGAC AGTGCCTCCT GGGCCCAGTT 540TGGCTGATGC TCTCATCAGT TCTCCGATTT TGGCTGGTGA AGGTGGTGCC ATGCTGGGTC 600TTGGTGCCAG TGACTTTGAA TTTGGAGTAG ATCCCAGTGC TGATCCTGAG CTGGCCTTGG 660CCCTTCGTGT ATCTATGGAA GAGCAGCGGC AGCGGCAGGA GGAGGAGGCC CGGCGGGCAG 720CTGCAGCTTC TGCTGCTGAG GCCGGGATTG CTACGACTGG GACTGAAGAC TCAGACGATG 780CCCTGCTGAA GATGACCATC AGCCAGCAAG AGTTTGGCCG CACTGGGCTT CCTGACCTAA 840GCAGTATGAC TGAGGAAGAG CAGATTGCTT ATGCCATGCA GATGTCCCTG CAGGGAGCAG 900AGTTTGGCCA GGCGGAATCA GCAGACATTG ATGCCAGCTC AGCTATGGAC ACATCTGAGC 960CAGCCAAGGA GGAGGATGAT TACGACGTGA TGCAGGACCC CGAGTTCCTT CAGAGTGTCC 1020TAGAGAACCT CCCAGGTGTG GATCCCAACA ATGAAGCCAT TCGAAATGCT ATGGGCTCCC 1080TGCCTCCCAG GCCACCAAGG ACGGCAAGAA GGACAAGAAG GAGGAAGACA AGAAGTGAGA 1140CTGGAGGGAA AGGGTAGCTG AGTCTGCTTA GGGGACTGCA TGGGAAGCAC GGAATATAGG 1200GTTAGATGTG TGTTATCTGT AACCATTACA GCCTAAATAA AGCTTGGCAA CTTTTAAAAA 1260AAAAAAAAAA AA 1272 ( 2 ) SEQ ID No.37

(ⅰ) sequence signature:

(A) length: 206 base pairs

(B) type: nucleic acid

(C) chain: strand

(D) topological classification: linearity, the sequence description of (ⅹ ⅰ) SEQ ID NO:37: CGGCACGAGA TGCCTACAGC TTCTCCCGGA AGATTACAGA GGCCATTGGC ATCATCAGCA 60AGATGATGTA TGAAAACACA ACTACAGTGG TGCAGGAGGT GATTGAATTC TTTGTGATGG 120TCTTCCAATT TGGGGTACCC CAGGCCCTGT TTGGGGTGCG CCGTATGCTG CCTCTCATCT 180GGTCTAAGGA GCCTGGTGTC CGGGAA 206, (2) information of SEQ ID No.38

(ⅰ) sequence signature:

(A) length: 341 base pairs

(B) type: nucleic acid

(C) chain: strand

(D) topological classification: the sequence description of linear (ⅹ ⅰ) SEQ ID NO:38: the information of TACTAAAAAT AAAAAATTAG CCGGGCGTAT TGGCGTGCGC CTGTAATCCC AGCTACTCAA 60GAGGCTGAGG CAGGAGAATC GCCTGAACCC AGAGGCGGAG GTTGTAGTGA GCCGAAATCA 120CACCATTGCA CTCCAGCTTG GGCAACAATA GCGAACCTCC ATCTCAAATT AAAAAAAAAA 180TGCCTACACG CTCTTTAAAA TGCAAGGCTT TCTCTTAAAT TAGCCTAACT GAACTGCGTT 240GAGCTGCTTC AACTTTGGAA TATATGTTTG CCAATCTCCT TGTTTTCTAA TGAATAAATG 300TTTTTATATA CTTTTAANGA GAGAAAAAAA ANAAACTCGA G 341 (2) SEQ ID No.39

(ⅰ) sequence signature:

(A) length: 293 base pairs

(B) type: nucleic acid

(C) chain: strand

(D) topological classification: the sequence description of linear (ⅹ ⅰ) SEQ ID NO:39: the information of CGGCACGAGC CCAGGCCCTG TTTGGGGTGC GCCGTATGCT GCCTCTCATC TGGTCTAAGG 60AGCCTGGTGT CCGGGAAGCC GTGCTTAATG CCTACCGCCA ACTCTACCTC AACCCCAAAG 120GGGACTCTGC CAGAGCCAAG GCCCAGGCTT TGATTCAGAA TCTCTCTCTG CTGCTAGTGG 180ATGCCTCGGT TGGGACCATT CAGTGTCTTG AGGAAATTCT CTGTGAGTTT GTGCAGAAGG 240ATGAGTTGAA ACCAGCAGTG ACCCAGCTGC TGTGGGAACC GGCCACCGAG AAA 293 (2) SEQ ID No.40

(ⅰ) sequence signature:

(A) length: 350 base pairs

(B) type: nucleic acid

(C) chain: strand

(D) topological classification: the sequence description of linear (ⅹ ⅰ) SEQ ID NO:40: the information of CGGCACGAGC TACCACCGCG TTCGGGTGTA GAATTTGGAA TCCCTGCGCC GCGTTAACAA 60TGAAGCAGAG TTCGAACGTG CCGGCTTTCC TCAGCAAGCT GTGGACGCTT GTGGAGGAAA 120CCCACACTAA CGAGTTCATC ACCTGGAGCC AGAATGGCCA AAGTTTTCTG GTCTTGGATG 180AGCAACGATT TGCAAAAGAA ATTCTTCCCA AATATTTCAA GCACAATAAT ATGGCAAGCT 240TTGTGAGGCA ACTGAATATG TATGGTTTCC GTAAAGTAAT ACATATCGAC TCTGGAATTG 300TTAAGCAAGA AAGAGATGGT CCTGTAGAAT TTCAGCATCC TTACTTCCAA 350 (2) SEQ ID No.41

(ⅰ) sequence signature:

(A) length: 377 base pairs

(B) type: nucleic acid

(C) chain: strand

(D) topological classification: the sequence description of linear (ⅹ ⅰ) SEQ ID NO:41: the information of TCCTAAAGCT TTCTCTGCTC CAGTTATTTT TATTAAATAT TTTTCACTTG GCTTATTTTT 60AAAACTGGGA ACATAAAGTG CCTGTATCTT GTAAAACTTC ATTTGTTTCT TTTGGTTCAG 120AGAAGTTCAT TTATGTTCAA AGACGTTTAT TCATGTTCAA CAGGAAAGAC AAAGTGTACG 180TGAATGCTCG CTGTCTGATA GGGTTCCAGC TCCATATATA TAGAAAGATC GGGGGTGGGA 240TGGGATGGAG TGAGCCCCAT CCAGTTAGTT GGACTAGTTT TAAATAAAGG TTTTCCGGTT 300TGTGTTTTTT TGAACCATAC TGTTTAGTAA AATAAATACA ATGAATGTTG NAAAAAAAAA 360AAAAAAAAAA ACTCGAG 377 (2) SEQ ID No.42

(ⅰ) sequence signature:

(A) length: 374 base pairs

(B) type: nucleic acid

(C) chain: strand

(D) topological classification: the sequence description of linear (ⅹ ⅰ) SEQ ID NO:42: the information of CGGCACGAGG CGCCACTTGC GAGCGCTGCA AGGGCGGCTT TGCGCCCGCT GAGAAGATCG 60TGAACAGTAA TGGGGAGCTG TACCATGAGC AGTGTTTCGT GTGCGCTCAG TGCTTCCAGC 120AGTTCCCAGA AGGACTCTTC TATGAGTTTG AAGGAAGAAA GTACTGTGAA CATGACTTTC 180AGATGCTCTT TGCCCCTTGC TGTCATCAGT GTGGTGAATT CATCATTGGC CGAGTTATCA 240AAGCCATGAA TAACAGCTGG CATCCGGAGT GCTTCCGCTG TGACCTCTGC CAGGAAGTTC 300TGGCAGATAT CGGGTTTGTC AAGAATGCTG GGAGACACCT GTGTCGCCCC TGTCATAATC 360GTGAGAAAGC CAGA 374 (2) SEQ ID No.43

(ⅰ) sequence signature:

(A) length: 492 base pairs

(B) type: nucleic acid

(C) chain: strand

( D ) ： ( ⅹⅰ ) SEQ ID NO:43：CTTTGCATTT TACAGTAAGA ATCAAAGTCC CTTCAGTGTG CCTTTGTCAG CTAATATGTG 60ACCAGCAATG ACAACCTTGG GAGTATTTAT TAAATATTAT GCTATGAATA TAGGCAACAC 120AGAACAGGGT TTGCAGTATA GCGTCTTGAT GCTAAATTCT CATATACCTC TACACGAGAA 180ATATGGAGGA GAAAAACAAG CATTTACATA TATTCTTCGT CACTTTGAAG ATGCATGACC 240TGAACTCGAC TGCTTGTGTT TGTTTACATA TCAGGCATAC CCAGGCATCT CCTGCAGCCA 300GAGGTTCCAT TGCTGTCTTT GCTCAGTCCT CTTTTAAAAT ATGAATTAGT GGACAGGCAC 360GGTGCCTCAC ACCTGTAATC CCAGCACTTT GGGAGGTCGA GGCAGGTGGA TCACGAGGTC 420AGGAGATCAA GACCATCCTG GCTACCACTG AAACCCCATC TCTACTACAA AAAAAAAAAA 480AAAAAACTCG AG 492 ( 2 ) SEQ ID No.44

(ⅰ) sequence signature:

(A) length: fifteen amino acid

(B) type: amino acid

(C) chain: strand

(D) topological classification: the sequence description of linear (ⅹ ⅰ) SEQ ID NO:44: the information of Ser Gln Ile Cys Glu Leu Val Ala His Glu Thr Ile Ser Phe Leu1 5 10 15 (2) SEQ ID No.45

(ⅰ) sequence signature:

(A) length: fifteen amino acid

(B) type: amino acid

(C) chain:

(D) topological classification: the sequence description of linear (ⅹ ⅰ) SEQ ID NO:45: the information of Xaa Xaa Xaa Xaa Xaa Ser Ile Leu Asp Glu Val Ile Arg Gly Thr1 5 10 15 (2) SEQ ID No.46

(ⅰ) sequence signature:

(A) length: 21 amino acid

(B) type: amino acid

(C) chain: strand

(D) topological classification: the sequence description of linear (ⅹ ⅰ) SEQ ID NO:46: Val Val Lys Thr Tyr Leu Ile Ser Ser Ile Pro Gln Gly Ala Phe Asn1 5 10 15Tyr Lys Tyr Thr Ala

The information of 20 (2) SEQ ID No.47

(ⅰ) sequence signature:

(A) length: 21 amino acid

(B) type: amino acid

(C) chain: strand

(D) topological classification: the sequence description of linear (ⅹ ⅰ) SEQ ID NO:47: Val Val Lys Thr Tyr Leu Ile Ser Ser Ile Pro Leu Gln Ala Phe Asn1 5 10 15Tyr Lys Tyr Thr Ala

The information of 20 (2) SEQ ID No.48

(ⅰ) sequence signature:

(A) length: fifteen amino acid

(B) type: amino acid

(C) chain: strand

(D) topological classification: the sequence description of linear (ⅹ ⅰ) SEQ ID NO:48: the information of Xaa Ala Lys Lys Phe Leu Asp Ala Glu His Lys Leu Asn Phe Ala1 5 10 15 (2) SEQ ID No.49

(ⅰ) sequence signature:

(A) length: fifteen amino acid

(B) type: amino acid

(C) chain: strand

(D) topological classification: the sequence description of linear (ⅹ ⅰ) SEQ ID NO:49: the information of Xaa Xaa Xaa Lys Ile Lys Lys Phe Ile Gln Glu Asn Ile Phe Gly1 5 10 15 (2) SEQ ID No.50

(ⅰ) sequence signature:

(A) length: 18 amino acid

(B) type: amino acid

(C) chain: strand

(D) topological classification: the sequence description of linear (ⅹ ⅰ) SEQ ID NO:50: the information of Xaa Lys Val Lys Val Gly Val Asn Gly Phe Gly Arg Ile Gly Arg Leu1 5 10 15Val Thr (2) SEQ ID No.51

(ⅰ) sequence signature:

(A) length: fifteen amino acid

(B) type: amino acid

(C) chain: strand

(D) topological classification: the sequence description of linear (ⅹ ⅰ) SEQ ID NO:51: the information of Xaa Tyr Gln Tyr Pro Ala Leu Thr Xaa Glu Gln Lys Lys Glu Leu1 5 10 15 (2) SEQ ID No.52

(ⅰ) sequence signature:

(A) length: 14 amino acid

(B) type: amino acid

(C) chain: strand

(D) topological classification: the sequence description of linear (ⅹ ⅰ) SEQ ID NO:52: the information of Xaa Pro Ala Val Tyr Phe Lys Xaa Xaa Phe Leu Asp Xaa Asp1 5 10 (2) SEQ ID No.53

(ⅰ) sequence signature:

(A) length: fifteen amino acid

(B) type: amino acid

(C) chain: strand

(D) topological classification: the sequence description of linear (ⅹ ⅰ) SEQ ID NO:53: the information of Xaa Pro Ala Val Tyr Phe Lys Glu Gln Phe Leu Asp Gly Asp Gly1 5 10 15 (2) SEQ ID No.54

(ⅰ) sequence signature:

(A) length: 18 amino acid

(B) type: amino acid

(C) chain: strand

(D) topological classification: the sequence description of linear (ⅹ ⅰ) SEQ ID NO:54: the information of Xaa Xaa Val Ala Val Leu Xaa Ala Ser Xaa Xaa Ile Gly Gln Pro Leu1 5 10 15Ser Leu (2) SEQ ID No.55

(ⅰ) sequence signature:

(A) length: fifteen amino acid

(B) type: amino acid

(C) chain: strand

(D) topological classification: the sequence description of linear (ⅹ ⅰ) SEQ ID NO:55: the information of Val Val Lys Thr Tyr Leu Ile Ser Xaa Ile Pro Leu Gln Gly Ala1 5 10 15 (2) SEQ ID No.56

(ⅰ) sequence signature:

(A) length: fifteen amino acid

(B) type: amino acid

(C) chain: strand

(D) topological classification: the sequence description of linear (ⅹ ⅰ) SEQ ID NO:56: the information of Xaa Xaa Lys Thr Tyr Leu Ile Ser Ser Ile Pro Leu Gln Gly Ala1 5 10 15 (2) SEQ ID No.57

(ⅰ) sequence signature:

(A) length: fifteen amino acid

(B) type: amino acid

(C) chain: strand

(D) topological classification: the sequence description of linear (ⅹ ⅰ) SEQ ID NO:57: Met Asp Ile Pro Gln Thr Lys Gln Asp Leu Glu Leu Pro Lys Leu1 5 10 15

Claims (31)

1. a peptide species, it comprises and has the SEQ of being selected from ID No.2, a kind of prostatein matter of 4,5,6,7 and 8 partial sequence or with this protein immunogenicity part of differentiated variant on conservative property is replaced and/or modified only.

2. a peptide species, its include a kind of prostatein matter or with this protein immunogenicity part of differentiated variant on conservative property is replaced and/or modified only, wherein this protein includes aminoacid sequence or its part of dna sequence encoding of the dna sequence dna of the complementary sequence that is selected from the sequence listed among SEQ ID No.11 and the 13-19, these sequences and the sequence of listing with SEQ ID No.11 and 13-19 or its complementary sequence hybridization under the moderate stringent condition.

3. dna molecular that contains the nucleotide sequence of coding claim 1 or 2 polypeptide.

4. expression vector that contains the dna molecular of claim 3.

5. expression vector transformed host cells with claim 4.

6. the host cell of claim 5, wherein this host cell is selected from intestinal bacteria, yeast and mammalian cell.

7. one kind includes the polypeptide of claim 1 or 2 and the pharmaceutical composition of physiologically acceptable carrier.

8. one kind includes the polypeptide of claim 1 or 2 and a kind of vaccine of nonspecific immune reaction toughener.

9. the vaccine of claim 8, wherein the nonspecific immune reaction toughener is a kind of adjuvant.

10. vaccine that contains a kind of dna molecular and a kind of nonspecific immune reaction toughener, this dna molecular include the nucleotide sequence of the polypeptide of coding claim 1 or 2.

11. the vaccine of claim 10, wherein the nonspecific immune reaction toughener is a kind of adjuvant.

12. a pharmaceutical composition that is used for prostate cancer therapy, it comprises a peptide species and a kind of physiologically acceptable carrier, and what this polypeptide contained a kind of prostatein has the SEQ of being selected from an ID No.1,3,20,21, the immunogenicity part of the partial sequence of 25-31 and 44-57.

13. a vaccine that is used for prostate cancer therapy, it includes a peptide species and a kind of nonspecific immune reaction toughener, and what this polypeptide included a kind of prostatein has the SEQ of a being selected from IDNo.1,3,20,21, the immunogenicity part of the partial sequence of 25-31 and 44-57.

14. the vaccine of claim 13, wherein the nonspecific immune reaction toughener is a kind of adjuvant.

15. the pharmaceutical composition according to claim 7, it is used to suppress the manufacturing of the medicine of prostate cancer development.

16. a vaccine according to Claim 8, it is used to suppress the manufacturing of medicine of the development of prostate cancer.

17. a method that detects prostate cancer among the patient comprises:

(a) available from patient's biological sample with can contact in conjunction with the binding reagents of the polypeptide of claim 1 or 2; And

(b) thus being attached to protein on the binding reagents or polypeptide in the test sample detects prostate cancer among the patient.

18. the method for claim 17, wherein binding reagents is a kind of monoclonal antibody.

19. the method for claim 17, wherein binding reagents is a kind of polyclonal antibody.

20. a method that monitors prostate cancer development among the patient comprises:

(a) available from patient's biological sample with can contact in conjunction with the binding reagents of the polypeptide of claim 1 or 2;

(b) be attached to the protein on the binding reagents or the amount of polypeptide in the test sample;

(c) repeating step (a) and (b); And

(d) comparison step (b) and (c) in the amount of detected polypeptide to monitor the development of prostate cancer among the patient.

21. a method that detects prostate cancer among the patient comprises:

(a) available from patient's biological sample with can contact in conjunction with the binding reagents of a peptide species, what this polypeptide contained a kind of prostatein matter has the SEQ of being selected from ID No.1,3,20,21, an immunogenicity part of the partial sequence of 25-31 and 44-57; And

(b) thus being attached to protein on the binding reagents or polypeptide in the test sample detects prostate cancer among the patient.

22. the method for claim 21, wherein binding reagents is a kind of monoclonal antibody.

23. the method for claim 21, wherein binding reagents is a kind of polyclonal antibody.

24. a method that monitors prostate cancer development among the patient comprises:

(a) available from patient's biological sample with can contact in conjunction with the binding reagents of a peptide species, what this polypeptide contained a kind of prostatein matter has the SEQ of being selected from ID No.1,3,20,21, an immunogenicity part of the partial sequence of 25-31 and 44-57;

(b) be attached to the protein on the binding reagents or the amount of polypeptide in the test sample;

(c) repeating step (a) and (b); And

(d) comparison step (b) and (c) in the amount of detected polypeptide to monitor the development of prostate cancer among the patient.

25. polypeptide bonded monoclonal antibody with claim 1 or 2.

26. the monoclonal antibody according to claim 25, it is used to suppress the manufacturing of medicine of the development of prostate cancer.

27. the monoclonal antibody of claim 26, monoclonal antibody wherein and the coupling mutually of a kind of therapeutical agent.

28. a method that detects prostate cancer among the patient comprises:

(a) contact with at least two Oligonucleolide primers in the polymerase chain reaction available from patient's biological sample, wherein at least one oligonucleotide primer is specific to and is selected from SEQ ID No.9-19, a kind of dna molecular of 22-24 and 32-43; And

(b) thus in the test sample when this Oligonucleolide primers exists the dna sequence dna of amplification detect prostate cancer.

29. the method for claim 28, wherein at least one Oligonucleolide primers comprises and is selected from SEQID No.9-19, a kind of dna molecular of 22-24 and 32-43 at least about 10 continuous nucleotides.

30. a method that detects prostate cancer among the patient comprises:

(a) be specific to available from patient's biological sample and at least one and be selected from SEQ ID No.9-19, the oligonucleotide probe of a kind of dna molecular of 22-24 and 32-43 contacts; And

(b) thus detect prostate cancer with the dna sequence dna of oligonucleotide probe hybridization in the test sample.

Be selected from SEQ ID No.9-19 31. the method for claim 30, probe wherein comprise, a kind of dna molecular of 22-24 and 32-43 at least about 15 continuous nucleotides.

Images