CN1995353A - Fungus pathogenic gene mgATG1 derived from rice blast and its uses - Google Patents

Abstract

The invention discloses a fungal pathogenic gene mgATG1 based on rice blast bacterium with nucleotide sequence or complementary nucleotide sequence as SEQ ID NO:1, wherein the cDNA sequence of gene mgATG1 possesses the nucleotide sequence as SEQ ID NO:2; the coded protein of gene mgATG1 possesses nucleotide sequence as SEQ ID NO:3; the promoter of gene mgATG1 possesses the nucleotide sequence as SEQ ID NO:4. The invention also provides an application of gene mgATG1 as target to sieve the antimycotic drug, which utilizes protein and promoter as target.

Description

Come from epiphyte pathogenic gene mgATG1 of Pyricularia oryzae and uses thereof

Technical field

The invention belongs to plant pathology, Pesticide Science and microbiological genetic engineering field, provide one derive from Pyricularia oryzae, nourish and grow, spore produces, appressorium forms with pathogenic in promotor and the nucleotide sequence of coding region and the aminoacid sequence of coded protein thereof of the new gene mgATG1 that plays an important role.Nucleotide sequence provided by the invention and aminoacid sequence can be used as in screening that target is applied to antifungal medicine and the design, and a certain section that also can utilize this nucleotide sequence is as probe or having among the clone of gene of certain homology with this sequence of utilizing that the antibody of this protein Preparation is applied to Pyricularia oryzae and other fungies.

Background technology

The rice blast that is caused by Pyricularia oryzae (Magnaporthe grisea) is the destructive disease on global a kind of paddy rice.The annual in the world rice yield that is caused by rice blast is lost between 10～30%.The almost annual popular outburst that Pyricularia oryzae is all arranged of China.Nearest 2005, Sichuan Province and Chongqing City were subjected to the most serious rice blast over 10 years, and rice blast takes place in 2,300,000 mu of rice fields that Sichuan Province has 20 cities, 127 counties, and the Chongqing City has more than 100 ten thousand mu of rice fields Pyricularia oryzae takes place; Current rice blast is popular rapidly, hazard rating is heavy, the morbidity rice varieties is many, it is serious that rice yield is influenced.Except paddy rice, Pyricularia oryzae also can infect other more than 50 kinds of grasses, also millet (Eleusine coracana), barley (Hordeum vulgare), wheat important farm crop such as (Triticum aestivum) is caused serious harm.Pyricularia oryzae also is the interactional model animals of a kind of research plant pathogenic fungi one host plant, have the total plant of many pathogenic fungies and cause a disease and infect circulation, comprise that spore generation, sprouting, appressorium form, infect bolt and form, invade pathogenic courses such as mycelial growth; Its pathogenic molecular mechanism is being studied by many developed countries, and expectation goes out the medicine target of novel pesticide screening test by this type of research and design, thereby develops, designs the novel anti fungi-medicine.

Include sexual stage and imperfect stage the life history of Pyricularia oryzae.Perfect stage produces ascus and thecaspore after by the strain mating of 2 different mating types.The mating type of Pyricularia oryzae is by mating gene M AT1-1 and MAT1-2 control.Imperfect stage is that the vegetative hyphae differentiation produces conidiophore and pyriform conidium.At nature, Pyricularia oryzae is mainly finished the life history by the imperfect stage, and conidium is the main intrusion source of plant.

After the Pyricularia oryzae asexual spore was fallen the rice leaf surface, infection processs began.After spore and water chance, the viscose glue in separate on the spore top discharges, and makes spore tightly stick to the rice leaf surface.The rice leaf surface has one deck wax cuticle, and this film has very big hydrophobicity, and viscose glue makes spore stick to this hydrophobic repellency surface.Adhere in back 2 hours, spore germination also produces germ tube.Germ tube stretches out and grows from an apical cell of spore usually.A kind of stickies is also secreted on the germ tube top, makes germ tube tightly stick to the plant leaf surface, prevents to be washed away by water droplet.In 4 hours, the germ tube growth stops, and the top crozier forms and expands and forms appressorium.The hardness of matrix is an important factor of germ tube differentiation, only forms appressorium at hard solid surface as magnaporthe grisea spore, and can not form appressorium at fluid surface or soft stromal surface.And the matrix hydrophobic surface also the effect of the wax by being similar to the rice leaf surface excite the formation of appressorium.Soon, the cell that produces germ tube carries out a mitotic division after spore germination.A nucleus is stayed in the spore, and another nucleus moves by germ tube, transfers to appressorium in the formation with the tenuigenin content of spore and germ tube.These tenuigenin contents comprise that the fat that is stored in the big vacuole of appressorium central authorities of growing drips.Then, between appressorium and germ tube, form barrier film.Along with the appressorium maturation, melanin deposition appears in the appressorium cell walls, finally forms the melanochrome layer.The melanochrome layer allows water molecules freely to pass through, but the material that is dissolved in water can not freely pass through, thereby allows appressorium set up and keep turgescence in the very big cell.After the melanochrome process of appressorium was finished, appressorium produced a narrow and small mycelia--infect bolt.Infecting bolt is accomplished penetrating owing to appressorium produces huge turgescence of rice leaf.Turgescence will stop appressorium to penetrate at artificial surface or rice leaf surface as long as reduce a little.Measuring result shows that the appressorium turgescence reaches 8.0Mpa, is equivalent to the pressure of 40 times of doughnuts.The effect of appressorium melanochrome layer has been to limit the perviousness of cell walls, helps the accumulation of permeate substance in the cell and the generation of turgescence.Glycerine is a kind of soluble substance, and the osmotic potential of generation is enough to make appressorium to produce huge turgescence.In the turgescence production process, glycerine surpasses 3M in cell inner accumulation concentration.

After the intrusion, infect bolt and be divided into and infect mycelia, growth and infect other cell in rice leaf rapidly.Invade after 72 hours, the pathogenic bacteria biomass has reached and has infected 10% of blade.After 5～7 days, differentiate a large amount of new conidiums on the conidiophore, and discharge from scab.The sporocyst malaria of these new formation takes contiguous plant to and begins new infection processs.In winter, Pyricularia oryzae is survived the winter on straw and paddy with mycelia and conidial form, finishes and infects circulation.

The pathogenic course of Pyricularia oryzae is the molecular process of a complexity.Cloned at present many genes relevant with rice blast bacterium pathogenicity, as: MPG1, CPKA, PMK1, MAGB, PLS1, SMO1, PDE1, MPS1, PTH11, CBP1, ICL1, BUF1, ALB1, ACR1, RSY1, HEX1.It is relevant that wherein most and appressorium form, as PMK1, MAGB, MAC1 etc.; The melanic formation of subparticipation and accumulation (ALB1, RSY1, BUF1 etc.) and glycerine synthetic relevant (ICL1 etc.); Minority relevant with penetrating of Pyricularia oryzae (MPS1, PLS1, PDE1 etc.).A kind of hydrophobin of Pyricularia oryzae germ tube excretory by the MPG1 genes encoding, participates in the mutual work of mycelia and paddy rice leaf table, for forming normal appressorium institute necessary (Talbot, 1999).The cAMP cyclase of the proteic alpha subunit of allos trimerization G of MAGB genes encoding and MAC1 coding also is to form appressorium necessary (Liu and Dean, 1997; Choiand Dean, 1997; Kulkarni and Dean, 2004).The PKA-c of catalytic subunit of the cAMP deopendent protein kinase A of CPKA genes encoding is that appressorium penetrates necessary (Mitchell ﹠amp; Hamer, 1995; Xu et al, 1997), and the PKA activity in the germ tube that is breaking up is for forming also very crucial (the Adachi ﹠amp of normal appressorium; Hamer1998).Equally, mitogen activated protein kinase (Mitogen-activated protein kinase, MAPK) gene PMK1 required (the Xu ﹠amp that is that appressorium forms; Hamer 1996).Melanochrome synthesizing series gene also proves appressorium and penetrates indispensable (Chumley and Valent, 1990; Motoyama etc., 1998).Found also that in various sudden change word banks some muton disappearances are pathogenic, as: a kind of proteic ABC1 gene (Urban etc. relevant with ATP carrier (ATP-bindingcassette transporters) encode, 1999), the PDE1 gene (Balhadere etc. of coding P-type ATP enzyme, 1999,2001) and the coding the proteic PLS1 gene of tetraspannin sample (Clergeot etc., 2001).However, the molecule mechanism of causing a disease of Pyricularia oryzae still is unclear.

The pathogenic gene of evaluation, clone plant pathogenic fungi, especially relevant with invasion procedure gene can be the target site that design, screening antifungal drug provide usefulness.Having proved at present the target spot of some medicines in comprising some fungies of Pyricularia oryzae, is a kind of very important Pyricularia oryzae sterilant as tricyclazole, and its effect is to suppress melanic synthetic, and target spot is three hydroxyl naphthalene reductase enzymes of Pyricularia oryzae; The action target spot of polypeptide sterilant sorphen A be mould demethylase (CYP51).Therefore, utilize the pathogenic gene that in the pathogenic course of Pyricularia oryzae, has critical function that Protocols in Molecular Biology is identified, the clone is new, can provide the drug target of usefulness for design, the new antifungal drug of screening.

Summary of the invention

The technical problem to be solved in the present invention provides a kind of new, mycelial growth to fungies such as Pyricularia oryzaes, generation of conidium, conidium and germinates, adheres to spore and form and infect that bolt forms and pathogenic gene mgATG1 that material impact is arranged and uses thereof.

In order to solve the problems of the technologies described above, the invention provides a kind of epiphyte pathogenic gene mgATG1 that comes from Pyricularia oryzae, the nucleotide sequence of this gene or its complementary nucleotide sequence are SEQ ID NO:1.

The present invention provides the cDNA sequence of said gene mgATG1 coding simultaneously, and this cDNA sequence has the nucleotide sequence shown in the SEQ IDNO:2.

The present invention provides said gene mgATG1 encoded protein matter simultaneously, and this protein has the aminoacid sequence shown in the SEQ ID NO:3.

The present invention provides the promotor of said gene mgATG1 simultaneously, and this promotor has the nucleotide sequence shown in the SEQ ID NO:4.

The present invention also provides the expression that utilizes said gene mgATG1 as target, uses in design and screening antifungal drug.

The present invention also provides and has utilized above-mentioned protein expression and modify as target, uses in design and screening antifungal drug.

The present invention also provides in conjunction with utilizing above-mentioned protein and promotor as target, uses in design and screening antifungal drug.

The protein sequence of gene mgATG1 of the present invention and this genes encoding is a probe with its full length nucleotide, separates the gene with identical function that comes from other pathogenic fungi by hybridization; Or, separate the gene that comes from other pathogenic fungi by PCR with identical function with the sequences Design primer; Or with its encoded protein matter, hybridization separates the albumen with identical function that comes from other pathogenic fungi.Above-mentioned fungi comprises: gibberella saubinetii (Gibberella zeae), botrytis cinerea (Botryotinia Fuckeliana), posadasis spheriforme (Coccidioides immitis), wheat glume blight bacterium (Phaeosphaeria nodorum), bean anthrax bacteria (Colletotrichum lindemuthianum) or corn smut (Ustilago maydis).

The alleged antifungal drug of the present invention comprises the medicine of resisting rice blast bacteria (Magnaporthe grisea), gibberella saubinetii (Gibberella zeae), botrytis cinerea (Botryotinia Fuckeliana), posadasis spheriforme (Coccidioidesimmitis), wheat glume blight bacterium (Phaeosphaeria nodorum), bean anthrax bacteria (Colletotrichumlindemuthianum) or corn smut (Ustilago maydis).

The present invention utilizes gene knockout and gene complementation to prove gene mgATG1 that the process that penetrates and the virulence of Pyricularia oryzae appressorium are had key effect in the Pyricularia oryzae.The promotor of this gene, the expression of proteins encoded and modification can be used as the drug target of novel pesticide design and screening.

Gene mgATG1 of the present invention clones from Pyricularia oryzae by the cDNA subtracted library.Detailed process comprises: the SSH subtracted library by screening Pyricularia oryzae cDNA obtains mycelia or adheres to the est sequence of differential expression in the spore.By RT-PCR checking goal gene at Pyricularia oryzae mycelia, spore, adhere to the situation that exists in the spore.From the Pyricularia oryzae genomic dna, obtain goal gene by the high-fidelity long range PCR.By PCR or with the est sequence of this gene is that probe is from checking of Pyricularia oryzae cDNA library and the complete cDNA that separates goal gene.The functional verification of goal gene and note are finished by knocking out with complementary recovery of goal gene.The action site of goal gene in Pyricularia oryzae determined in the position of cell by the fusion rotein of goal gene and reporter gene GFP.The mensuration of gene promoter is that promotor is connected with reporter gene GFP, and GFP carries out in the expression of each etap of Pyricularia oryzae under promotor instructs by observing.

Pyricularia oryzae SSH subtracted library involved in the present invention is meant and utilizes SSH technology (Suppression subtractivehybridization, SSH) Pyricularia oryzae cDNA library that make up, that contain some independent clonings.RT-PCR involved in the present invention analyzes and is meant according to cDNA sequences Design primer, the cDNA that genetically deficient muton and the complementary mRNA reverse transcription that recovers muton are formed carries out pcr amplification, verify respectively genetically deficient muton and gene complementation recover genetically deficient in the muton and recover after situations such as transcript.

Goal gene clone involved in the present invention is meant the est sequence that obtains according in the SSH library, screens the cDNA clone with this designing probe from the cDNA library, and the cDNA clone who obtains is checked order; And utilize dna sequence dna in the cDNA sequence alignment genome database, and design the dna sequence dna of primer with this from Pyricularia oryzae genomic dna amplification acquisition gene, be cloned in the T carrier, check order; Utilize its transcription sequence of programanalysis according to the dna sequence dna that obtains, and design from Pyricularia oryzae cDNA library, the increase cDNA sequence of acquisition gene of primer, be connected in the T carrier, carry out sequencing analysis with this.

Shouthern hybridization analysis involved in the present invention is meant that the dna sequence dna that utilizes the gene flank is as probe, gene knockout transformant and the gene recovery transformant that is obtained carried out Southern hybridization respectively, knock out the genetically deficient situation of transformant and the insertion copy number of knockout carrier with analyzing gene, and the complementary transformant gene that recovers of analyzing gene inserts situations such as copy number.

The function of goal gene involved in the present invention is determined knocking out and recover to carry out by mgATG1.The process of knocking out of concrete mgATG1 gene comprises the structure of knockout carrier, changes knockout carrier over to Pyricularia oryzae, obtains the gene knockout muton; The complementary recovery process of the gene knockout muton of mgATG1 comprises the structure of complementary carrier, and complementary carrier quiding gene knocks out the muton bacterial strain, obtains the transformant that gene recovers.The structure of gene knockout carrier is meant that the section of DNA fragment that will wait to knock out two flanks of gene is connected to the both sides of the hygromycin gene that contains the hygromycin gene carrier separately.The structure of gene complementation carrier be meant with contain target gene total length dna fragmentation with have a carrier that is different from the selection markers (being the Glufosinate ammonium resistant gene among the present invention) of hygromycin gene and be connected.The preferred method that in the present invention foreign vector is imported Pyricularia oryzae is a protoplast transformation, and the experimenter also can select other method such as ATMA method, particle bombardment etc. for use according to the situation of oneself.Among the present invention, the Pyricularia oryzae bacterial strain that gene knockout carrier imported is wild type strain Guy-11, the experimenter also can select other Pyricularia oryzae wild type strain for use, and genetically deficient makes the phenotype of muton be different from the variation of original wild-type, to the pathogenic forfeiture of plants such as paddy rice.Among the present invention, the bacterial strain that the gene complementation carrier is imported is the resulting genetically deficient muton of the present invention, and the ectopic integration of complementary carrier in this genetically deficient muton genome makes the phenotype of this muton recover normal.

The cDNA clone process of mgATG1 provided by the present invention comprises: screening contains the cDNA clone of target gene mgATG1 from the SSH subtracted library, obtain its est sequence, utilize its est sequence from the Pyricularia oryzae genome database, to obtain corresponding section of DNA sequence, dna fragmentation at the target gene that from the Pyricularia oryzae genomic dna, increases according to its sequences Design primer, order-checking obtains its dna sequence dna, utilizes GenScan (http://genes.mit.edu/genscan.html) to carry out the prediction of promotor, coding region and tailing point.According to the transcript sequences Design primer of prediction, and by the PCR acquisition of increasing from Pyricularia oryzae cDNA library, its sequence is shown in SEQ IDNO:2.The present invention also provides mgATG1 encoded protein matter, and it has shown in SEQ ID NO:3 aminoacid sequence or has aminoacid sequence sequence identity and that have identity function more than 35% with the sequence shown in the SEQ IDNO:2.This albumen can come from Pyricularia oryzae, also can come from other pathogenic fungi.The invention provides gibberella saubinetii (Gibberella zeae), botrytis cinerea (Botryotinia Fuckeliana), posadasis spheriforme (Coccidioidesimmitis), wheat glume blight bacterium (Phaeosphaeria nodorum), bean anthrax bacteria (Colletotrichumlindemuthianum), corn smut (Ustilago maydis) in have the aminoacid sequence that has the homologous protein of 35% above consensus amino acid sequence with mgATG1 albumen.

Gene involved in the present invention is because the genetically deficient sudden change that homologous recombination causes causes the decline of Pyricularia oryzae sporulation quantity, spore germination delay, the reduction of appressorium turgescence and invades the bolt rate of formation and reduce, and the virulence on susceptible rice varieties lacks.Therefore, the most important purposes of the present invention is: use above-mentioned achievement, design and screening can destroy the compound of the expression of this expression of gene, shearing and proteins encoded thereof, or the compound that can modify this proteic aminoacid sequence of design and screening, thereby the antifungal drug that exploitation makes new advances.In addition, purposes of the present invention comprises that also the DNA that utilizes this gene or cDNA sequence separate with this gene as probe and have the sequence of certain sequence homology in other fungi.

With the expression of above-mentioned institute clone gene proteins encoded, be modified to target; design, screening novel anti fungi-medicine; perhaps to have shown in SEQ ID No:3 sequence or to have the aminoacid sequence design polypeptide in arbitrary zone in the albumen 35% above sequence identity and that have identity function with SEQ ID No:3; and preparation antibody is used for detecting proteic expression under the compound treatment situation, all belongs within protection scope of the present invention.Promotor with above-mentioned institute clone gene is a target, and design and screening novel anti fungi-medicine also belong within protection scope of the present invention.

Therefore, the present invention has the following advantages:

1, appressorium is the important structure that Pyricularia oryzae is invaded host plant, the appressorium turgescence is the main mechanical forces that Pyricularia oryzae penetrates the host plant horny layer of epidermis, the mgATG1 gene participates in the generation of Pyricularia oryzae appressorium turgescence, mgATG1 gene and Pyricularia oryzae pathogenic closely related, mgATG1 inactivation or disappearance, cause Pyricularia oryzae appressorium turgescence to descend pathogenic forfeiture.Therefore the mgATG1 gene acts on great in the rice blast pathogenic process.

2, mgATG1 provides special drug effect target for developing novel antifungal medicines, at the target medicine target of mgATG1 as drug development, by a large amount of compounds are screened, can find the material that can work with the medicine target, these materials can be used as the prevention fungal infection.Because the specificity of medicine target, the antifungal drug that screens is generally less for the toxicity of animal and plant.

Description of drawings

Fig. 1 is wild type strain Guy11 and mgATG1 genetically deficient muton s197-4 and the pathogenic comparison diagram of the complementary transformant HB24 of mgATG1 on paddy rice susceptible variety CO39.Guy11 is a wild type strain, and s197-4 is a mgATG1 genetically deficient muton, and HB24 recovers transformant for the mgATG1 gene complementation, and gelatin is the gelatin contrast solution of 0.2% (w/v).The spore concentration of spray inoculation is 1 * 10 ⁵/ ml inoculates back 7 days and takes pictures.

Fig. 2 is that the complementary transformant HB24 of wild type strain Guy11 and mgATG1 genetically deficient muton s197-4 and mgATG1 invades bolt formation comparison diagram on the barley epidermis.A is wild type strain Guy11, and B is mgATG1 genetically deficient muton s197-4, and C recovers transformant HB24 for the mgATG1 gene complementation; 24h, 48h, 72h, 96h are inoculation back photo opporunity.The spot-inoculated spore concentration that exsomatizes is 1 * 10 ⁵/ ml.

Fig. 3 is that gene knockout carrier makes up synoptic diagram.

Fig. 4 is a gene complementation vector construction synoptic diagram.

Fig. 5 is gene knockout position and process synoptic diagram.

Fig. 6 is the big or small comparison diagram of the appressorium turgescence that forms of the complementary transformant HB24 of wild type strain Guy11 and mgATG1 genetically deficient muton s197-4 and mgATG1 (appressorium subside rate).Last figure is the cell of spore inoculating 24h postadhesion born of the same parents in the different glycerol concentration solution rate of subsiding, and figure below is the cell of spore inoculating 48h postadhesion born of the same parents in the different glycerol concentration solution rate of subsiding.

Fig. 7 is wild type strain Guy11 and the mgATG1 genetically deficient muton s197-4 comparison diagram that autophagic vacuole forms in the nitrogen stress substratum.Upper left (A) and lower-left (C) figure are the electromicroscopic photograph that wild type strain Guy11 bacterial strain autophagic vacuole forms, and upper right (B) and bottom right (D) figure are the electromicroscopic photograph that mgATG1 genetically deficient muton s197-4 bacterial strain autophagic vacuole forms.

Fig. 8 is the glycogenosome comparison diagram in the spore that forms of the complementary transformant HB24 of wild type strain Guy11 and mgATG1 genetically deficient muton s197-4 and mgATG1.Last figure (A) is wild type strain Guy11, and middle figure (B) is mgATG1 genetically deficient muton s197-4, and figure below (C) is the complementary transformant HB24 of mgATG1.

Fig. 9 is the tenuigenin location map of mgATG1 gene coded protein.The mgATG1-GFP fusion rotein is distributed in the tenuigenin, and the left side is to take under the bright field, and the right side is to take under the dark field.Upper left (A) is the spore photo under the common light microscopic, upper right (B) is the spore photo under the fluorescent microscope, (C) is the mycelia photo under the common light microscopic in the left side, (D) is the mycelia photo under the fluorescent microscope in the right side, lower-left (E) is the appressorium photo under the common light microscopic, and bottom right (F) is the appressorium photo under the fluorescent microscope.

Figure 10 is the sporulation quantity comparison diagram of wild type strain Guy11 and mgATG1 genetically deficient muton s197-4 and the complementary transformant HB24 of mgATG1.

Figure 11 is wild type strain Guy11 and mgATG1 genetically deficient muton s197-4 and the speed of growth comparison diagram of the complementary transformant HB24 of mgATG1 on perfect medium, nitrogen stress substratum, scarce charcoal substratum.CM is a perfect medium, and MM-N is the nitrogen stress substratum, and MM-C is for lacking the charcoal substratum.

Figure 12 is the spore germination rate comparison diagram of wild type strain Guy11 and mgATG1 genetically deficient muton s197-4 and the complementary transformant HB24 of mgATG1.

Figure 13 is the expression figure of mgATG1 gene in spore and mycelia.Left side (A) is taken down for the bright field, and right side (B) takes down for dark field.

Figure 14 is the proteic sequence comparison diagram of ATG1 in several pathogenic fungies.Illustrate: mgATG1, gzATG1, bfATG1, ciATG1, pnATG1, clATG1, umATG1 is respectively from Pyricularia oryzae (Magnaporthegrisea), gibberella saubinetii (Gibberella zeae), botrytis cinerea (Botryotinia Fuckeliana), posadasis spheriforme (Coccidioides immitis), wheat glume blight bacterium (Phaeosphaeria nodorum), bean anthrax bacteria (Colletotrichum lindemuthianum), the ATG1 albumen of corn smut (Ustilago maydis).

Embodiment

Below in conjunction with accompanying drawing the specific embodiment of the present invention is described in further detail.

Embodiment 1: the clone of the gene relevant with rice blast bacterium pathogenicity:

1, SSH library screening: the difference that Pyricularia oryzae appressorium cDNA and sporogenous hyphae cDNA difference subtract the back acquisition subtracts cDNA, be cloned into the T carrier, order-checking obtains the cDNA sequence, the sequence and the Pyricularia oryzae database that obtain are compared, determine the difference expression gene of acquisition, and verify the differential expression specificity with RT-PCR.

2, the clone of mgATG1 cDNA: the appressorium cDNA library of the Pyricularia oryzae Guy11 bacterial strain of existence-20 ℃ of going bail for, gene cDNA sequence design synthetic pcr primer thing according to prediction, amplification comprises whole gene coding region at interior gene cDNA fragment, be cloned into the T carrier, order-checking obtains the sequence of mgATG1 cDNA, and its sequence is shown in SEQ ID NO:2.

3, the clone of mgATG1 genomic dna: the genomic dna of the Pyricularia oryzae Guy11 bacterial strain of existence-20 ℃ of going bail for, gene DNA sequence design synthetic pcr primer thing according to prediction, amplification comprises the dna fragmentation of whole full-length gene, be cloned into the T carrier, order-checking obtains the sequence of mgATG1 genomic dna, and its sequence is shown in SEQ ID NO:1.

The acquisition and the evaluation of embodiment 2:mgATG1 genetically deficient muton

1, the structure of gene knockout carrier: the dna fragmentation that is about 500～2000bp in mgATG1 upstream region of gene and downstream is inserted into the both sides of hygromycin gene in the carrier that contains hygromycin gene respectively, as Fig. 3 and shown in Figure 5.Specific as follows: the method for employing high-fidelity PCR is the dna fragmentation that is about 500～2000bp in amplification mgATG1 upstream region of gene and downstream from Pyricularia oryzae DNA genome respectively, is connected to earlier on the T carrier; The upstream dna fragmentation downcuts from the T carrier with restriction enzyme Xhol and Sall then, is inserted on the same restriction enzyme site of the pBSHPH1 carrier that contains hygromycin gene; Then the downstream DNA fragment is also downcut from the T carrier with restriction enzyme Hindlll and Xbal from the T carrier, is inserted on the Hindlll and Xbal site of the above-mentioned pBSHPH1 carrier that has inserted the upstream dna fragmentation, promptly obtains the mgATG1 gene knockout carrier.The building process that contains the pBSHPH1 carrier of hygromycin gene: adopt earlier the method for the high-fidelity PCR hygromycin gene (HPH or HygBr) that from the pCB1003 plasmid, increases, be connected on the T carrier; Downcut with restriction enzyme Hindlll and Sall then, and be inserted in the same restriction enzyme site of pBS plasmid, promptly obtain the pBSHPH1 carrier.

2, the preparation of Pyricularia oryzae protoplasma carrier and DNA transform: the Pyricularia oryzae bacterial strain is earlier growth on the CM flat board about 6 days.Cut the bacterium colony of two about 3cm of diameter, place 150mL CM liquid nutrient medium, it is smashed to pieces with triturator; Evenly divide then to install in two bottles of 200mL CM liquid nutrient mediums, 28 ℃, 125rpm cultivated 48 hours.Four layers of filtered through gauze are collected mycelia, with aseptic water washing twice, moisture are pressed dry.Mycelia in enzymolysis solution (Glucanex, enzyme liquid concentration 7.5mg/mL, 0.7M NaCl solution) enzymolysis 2-3hr 30 ℃, 80rpm.Take out enzymolysis solution, three metafiltration paper filter, and residue is removed in 0.7M NaCL washing, and filtrate collection is in the centrifuge tube of 50mL.4 ℃, 3000rpm, centrifugal 10min.With 10-20mL STC (1.2M Sorbitol, 10mM Tris-HCL PH7.5,20mM CaCL2) suspension precipitation.4 ℃, 3000rpm, centrifugal 10min; Repeat twice.Get an amount of STC suspension precipitation, making final concentration is 0.5～1.0 * 10 ⁸Individual/mL.Each 50ml centrifuge tube packing 150 μ l protoplastis adds 2 μ g linearizing DNA, places 25min under the room temperature.Dropwise add 1mL PTC (60%PEG4000,10mM Tris-HCL PH7.5,20mMCaCL ₂), mixing, room temperature is placed 25min.Slowly add 5mL OCM liquid nutrient medium (adding 1.2M Sorbitol/1M Sucrose in the CM liquid nutrient medium), shake up gently, place 28 ℃, 100rpm shakes and spends the night.Add the solid OCM Top substratum that contains 200 μ g/mL Totomycin in the protoplastis of every pipe incubated overnight and (add 20%Sucrose in the GM liquid nutrient medium, agar powder 1g), mixing, pour into and be covered with OCM Bottom substratum and (add 20%Sucrose in the CM liquid nutrient medium, agar powder 1.5g) flat board, three flat boards in every pipe shop.Dark, 28 ℃, to cultivate about an about week, the picking transformant is inoculated on the solid CM that contains 200 μ g/mL Totomycin.

3, the evaluation of mgATG1 genetically deficient muton: extract transformant DNA, with method validation transformants such as PCR, Southern hybridization with hygromycin resistance.

The complementation of embodiment 3, mgATG1 genetically deficient muton recovers:

The full length DNA sequence of mgATG1 gene is connected in the carrier that has the Glufosinate ammonium resistant gene, as shown in Figure 4.Be specially: adopt high-fidelity long segment PCR method (LD PCR) from the Pyricularia oryzae genomic dna, to increase and obtain containing the dna fragmentation in mgATG1 full length gene coding region, promoter region and zone, terminator, be inserted into earlier in the pCR-XL-TOPO carrier, and then from the TOPO carrier, downcut this dna fragmentation with restriction enzyme EcoRI, be inserted into the EcoRI site of the pBARKS1 carrier that contains the Glufosinate ammonium resistant gene, the complementation that promptly is built into the mgATG1 gene recovers carrier---pBARATG1 carrier.Before the protoplasma that carries out DNA transformed, the pBARATG1 carrier was being used restriction enzyme Notl linearization for enzyme restriction.

Then according to the method for embodiment 2, the complementary carrier of linearizing this mgATG1 gene is transformed mgATG1 genetically deficient bacterial strain protoplastis.The transformant that picking has the Glufosinate ammonium resistance is inoculated on the solid CM that contains Glufosinate ammonium.Extraction has the transformant DNA and the RNA of Glufosinate ammonium resistance, with method validation transformants such as RT-PCR, Southern hybridization.

The changing condition that has compared phenotype after gene recovers by the following method: relatively the sporulation quantity of the sub and complementary recovery transformant of mgATG1 transgenation changes (see figure 10) on perfect medium; MgATG1 transgenation and the complementary sprouting variation (seeing Figure 12) that recovers the transformant spore have been compared; The turgescence of adhering to spore that has compared mgATG1 transgenation and the formation of complementary recovery transformant changes (see figure 6); The mycelia autophagic vacuole that has compared mgATG1 transgenation and the formation of complementary recovery transformant forms the situation (see figure 7); The glycogen content that has compared the spore of mgATG1 transgenation and the formation of complementary recovery transformant changes (see figure 8); On perfect medium, nitrogen stress substratum, scarce carbon substratum, compare mgATG1 transgenation and the complementary growth change (seeing Figure 11) of recovering transformant; Fig. 1 is seen in the pathogenic recovery of complementary transformant.

The pathogenic mensuration of embodiment 4:mgATG1 genetically deficient muton and complementary muton:

1, spray inoculation: barley or paddy rice CO39 seed kind are cultivated 8 days (barley) or 14 days (the paddy rice CO39 plant of 3～4 leaf phases) in flowerpot earth.Wash the spore of getting Pyricularia oryzae wild type strain, muton and complementary transformation from the CM flat board of cultivating 12 days, spore concentration is diluted to 1 * 10 with the gelatin of 0.2% (wt/v) ⁵/ ml.With miniature airbrush with the spore suspension spray inoculation to the plant leaf surface.Simultaneously, with 0.2% gelatin as the negative control spray inoculation.The inoculation plant is placed in the moist chest, preserves moisture in 25 ℃ of dark surrounds and cultivates 24 hours (barley) or 48 hours (paddy rice).Inoculate plant then and put into and continue to be cultured to the 4th day (barley) or the 7th day, the 14th day (paddy rice) in the growth cabinet (25 ℃, 80% above humidity, 12 hours periodicity of illuminations), the plant illness is fully shown.The rice blast symptom of Taking Pictures recording plant leaf, and according to (1986) reported method such as Bonman records with estimate the disease serious degree of plant.Simultaneously, tried to select the plant a slice the most serious infected blade, the Pyricularia oryzae scab on the record blade in one section zone of 5cm length from every strain.The results are shown in Figure 1.

2, in vitro inoculation: the barley seed kind was cultivated 7～10 days in flowerpot earth.Wash the spore of getting Pyricularia oryzae wild type strain, muton and complementary transformation from the CM flat board of cultivating 12 days, being diluted to spore concentration is 5 * 10 ⁴/ ml or 1 * 10 ⁵/ ml.Spore liquid 20 μ l one dropping point is inoculated in the barley leaves upper surface that exsomatizes.In growth cabinet, preserve moisture illumination cultivation (illumination/dark, 12h:12h), 25 ℃.Cultivating 24h respectively, 48h behind 72h and the 96h, takes out partial blade and observes.Observe scab earlier and form situation, and Taking Pictures recording.Clip part scab is put into methanol solution and is fixed and slough chlorophyll then.The scab blade is fixed 24 hours (room temperature) in methanol solution after, (lactophenol oil: 95% ethanol=1: 2) room temperature is fixed 1 hour to move into the fixing transparent liquid of lactophenol oil.The lactophenol oil formula is: lactic acid 20ml, phenol 20ml, 20% glycerine 40ml, distilled water 20ml.The scab blade is used tongue phenol indigo plant/aniline blue (0.01% tongue phenol indigo plant/0.06% aniline blue, lactophenol oil) dyeing 5～10 minutes again.The dyeing blade decolours in lactophenol oil.Examine under a microscope the scab development, the appressorium penetration coefficient of statistics 24h and 48h and the growth of intrusion mycelia and the production of secondary spore, and Taking Pictures recording.The results are shown in Figure 2.

The Subcellular Localization of embodiment 5:mgATG1:

The complete coding region sequence of GFP gene is connected to the downstream of the mgATG1 gene coding region of not being with terminator codon, is built into fusion gene and is connected in the carrier that has hygromycin gene.According to the method for embodiment 2, linearizing this carrier is transformed wild-type Guy11 bacterial strain protoplastis.Picking has the transformant of hygromycin resistance and observe the distribution of mgATG1-GFP fusion rotein in the Pyricularia oryzae cell under fluorescent microscope.The result shows that mgATG1 is positioned in the tenuigenin.The results are shown in Figure 9.

Defining and expression analysis of embodiment 6:mgATG1 promotor:

After the complete coding region sequence of GFP gene is connected to the mgATG1 gene coding region upstream section of DNA sequence of (not containing coding region sequence), is built into fusion gene and is connected in the carrier that has hygromycin gene.According to the method for embodiment 2, linearizing this carrier is transformed wild-type Guy11 bacterial strain protoplastis.GFP albumen under the mgATG1 promotor of the transformant that picking has a hygromycin resistance observing under fluorescent microscope instructs the results are shown in Figure 13 at the expression of Pyricularia oryzae different developmental phases.The mgATG1 promoter sequence is seen shown in the SEQ ID NO:4.

Embodiment 7: the proteic bioinformatic analysis of fungi ATG1:

For whether clear and definite ATG1 albumen is guarded in other pathogenic fungies, at following pathogenic fungi: gibberella saubinetii (Gibberella zeae), botrytis cinerea (Botryotinia Fuckeliana), posadasis spheriforme (Coccidioidesimmitis), wheat glume blight bacterium (Phaeosphaeria nodorum), bean anthrax bacteria (Colletotrichumlindemuthianum), corn smut (Ustilago maydis, the proteic aminoacid sequence of mgATG1 is carried out the Blastp retrieval to the protein sequences of these fungies, obtain the aminoacid sequence of the homologous protein of above-mentioned pathogenic fungi, its sequence is respectively as SEQ ID NO:5,6,7,8, shown in 9 and 10; Be specially: SEQ ID NO:5 represents gzATG1 Gibberellazeae gibberella saubinetii, SEQ ID NO:6 represents bfATG1 Botryotinia Fuckeliana botrytis cinerea, SEQ ID NO:7 represents ciATG1 Coccidioides immitis posadasis spheriforme, SEQ ID NO:8 represents pnATG1Phaeosphaeria nodorum wheat glume blight bacterium, SEQ ID NO:9 represents clATG1 Colletotrichumlindemuthianum bean anthrax bacteria, and SEQ ID NO:10 represents umATG1 Ustilago maydis corn smut.And with these albumen difference called afters gzATG1, bfATG1, ciATG1, pnATG1, clATG1 and umATG1.Sequential analysis shows that the ATG1 albumen of above-mentioned fungi reaches 59%, 51%, 50%, 47%, 44%, 36% respectively with the proteic consistence of mgATG1 on aminoacid sequence.Utilize PAUP ^*4.0b10 the proteic homologous relationship figure of ATG1 of the above-mentioned fungi that program is drawn sees Figure 14.

Embodiment 8: utilize the proteic expression of mgATG1 or modify as target the screening antifungal medicament.

Pyricularia oryzae is cultivated in the liquid perfect medium in a large number, be divided into some aliquots after collecting mycelia, adding compound to be screened or drug candidate in each part continues to cultivate a few hours at perfect medium, in the nitrogen stress substratum, (also add compound or drug candidate to be screened simultaneously) then and continue inducing culture about 2 hours, get mycelia is observed mycelia under phase microscope autophagic vacuole formation situation, perhaps mycelia utilizes conventional electron microscope slice production method to make the formation situation of observing autophagic vacuole behind the ultrathin section(ing) under electron microscope.If mgATG1 albumen takes place to be modified incident by drug candidate and lose activity, then mycelia does not form autophagic vacuole.The compound that obtains utilizes the method for embodiment 4 again, measures the wild-type Pyricularia oryzae and under the condition that this compound exists whether the pathogenic of paddy rice is weakened, and further determines the anti-mycotic efficiency of compound.

Embodiment 9: the expression that utilizes mgATG1 is screened antifungal medicament as target.

Pyricularia oryzae is cultivated in the liquid perfect medium in a large number, be divided into some aliquots after collecting mycelia, adding compound to be screened or drug candidate in each part continues to cultivate a few hours at perfect medium, in the nitrogen stress substratum, (also add compound or drug candidate to be screened simultaneously) then and continue inducing culture about 2 hours, extract the RNA of mycelia, adopt the expression situation of the method detection mgATG1 of real-time quantitative PCR.If the expression of mgATG1 is suppressed by drug candidate, then there is not the transcript of mgATG1 in the hyphal cell.The compound that obtains utilizes the method for embodiment 4 again, measures the wild-type Pyricularia oryzae and under the condition that this compound exists whether the pathogenic of paddy rice is weakened, and further determines the anti-mycotic efficiency of compound.

Embodiment 10: the promotor of utilizing mgATG1 is screened antifungal medicament as target.

The promotor of mgATG1 and fluorescin GFP are built into the fusion rotein carrier, or the promotor of mgATG1 is built into the fusion rotein carrier with fluorescin GFP and mgATG1 albumen, method by embodiment 2 is transferred in the Pyricularia oryzae, then the Pyricularia oryzae transformant bacterial strain of expressing at the GFP under the promoter regulation of mgATG1 is cultivated in the liquid perfect medium in a large number, be divided into some aliquots after collecting mycelia, add compound to be screened or drug candidate in each part and continue to cultivate a few hours to a couple of days, get mycelia is observed the GFP of mycelia under fluorescent microscope green fluorescence expression at perfect medium.If the combined thing of the promotor of mgATG1 suppresses and loses activity, thereby then GFP albumen is not expressed mycelia and is lost green fluorescence.The compound that obtains utilizes the method for embodiment 4 again, measures the wild-type Pyricularia oryzae and under the condition that this compound exists whether the pathogenic of paddy rice is weakened, and further determines the anti-mycotic efficiency of compound.

At last, it is also to be noted that what more than enumerate only is several specific embodiments of the present invention.Obviously, the invention is not restricted to above embodiment, many distortion can also be arranged.All distortion that those of ordinary skill in the art can directly derive or associate from content disclosed by the invention all should be thought protection scope of the present invention.

Sequence table

SEQ?ID?NO：1

aagaagatgg?aggggacgct?aaatggtacc?gatctggcta?aagcctgcca?agcagccacc?60

aactcgtgac?agaggaacag?aatggcatct?gtgcgaagtc?tcttggttgc?ttctcaacga?120

atgcagtcgt?tcatcaggcg?ttctatttgc?tgcctcacat?aggcgagttt?gggtaggcag?180

cttaccttat?ctgtcgtgtt?gtcatttcgc?aaggaaagtc?gcaacaggtt?gtcatggatc?240

gtctaatcaa?ttacgtctat?cagttccaag?ctatcatcat?ctcaacctga?ctaagtgtgt?300

agtacccatc?tatgtatctt?tgggcatggc?atgcctaccg?cgtatcacac?cctcgaaatc?360

cacagtgcca?caattatcat?ccccaactat?gcagggcact?gtcgtttctc?atttacccat?420

cttatctgct?ctctgcgtcg?taaatctccg?tctttattgc?ctagtgcctt?tacacttctt?480

ttattagaca?ccgcgttggg?ggttctggaa?taacttactc?cgtacaatag?atagctttct?540

ttttgttctt?tctttctttc?tttttttttt?tttttttaaa?aaaaaaagat?aggaaaaaga?600

aagaaagaaa?gaaaagactg?cacttaatat?ctgaaatctc?agttcagtcc?aaggcaaggt?660

ctctaaaggt?acctacctag?ataggtacct?accaggtacg?taccttacgt?acctcactga?720

ctccgccaaa?ccccacactg?gtgtcgtcaa?cccaagttcc?gatcacgcac?cagtctttgc?780

ccattgctat?ggagctcacc?agacctgaca?cgccggggac?agtgcaggcg?tcacggttga?840

cgccaaatta?ggcatcccgg?acatgaagtt?tatgttacgg?tgttgtaaat?aattattaca?900

ctgacacgaa?tgcagccttt?tgacagaagt?caaccttcct?aggccacaaa?actcttatta?960

ctctgcttgc?tcggtcctag?atctggacgt?acagtaggta?attggtaggt?acctagttag?1020

gtaggtaggt?tactgtattg?taatggaggt?gaggtgatga?ccgtacagta?ccggtggtga?1080

ttgtgcagcc?agttcagtgg?catccgcccg?cccagtgccg?ttaaaaaaaa?aaaaaaccac?1140

ccgctcatta?gaccgcccac?ccccagctgc?cacctgccca?cgtcttacac?aaactccgaa?1200

aaacgaacga?cttaccctgt?ttgttaccac?tttcaattgc?cttatcttct?catcaaagcc?1260

gtcacgagtt?ggggatcaaa?ctcgaaaaaa?atcgaggcct?tcaaagacac?gtcttgaaag?1320

aggcttggcc?catttgcgaa?tagaaaaagg?tcggtatgtc?ttgtggtgac?gtcgttttcg?1380

ctttcaacct?gtcgccccct?taaactattg?tttacttctc?ggtcgcaggc?tatgcagcgg?1440

agcgctaggc?gcgtggaacc?ccctgccaca?agtcacttgc?gctccacatc?caatccccac?1500

acatgatcca?gaacctcgtc?gtgaatcctg?ggcctatcgg?agccgtgcca?gagaccaggc?1560

ggcggaccag?ttcctaccat?tcgtgacctt?gcctgcctgc?ctccccggcc?tagcgatcca?1620

ggcaccttgt?tgactgcttt?gaactgtgct?tgctcatcat?cacccaactt?ttctcttacc?1680

tatctttttt?ctgtttttcc?ctcccgagtc?ttttcatccc?atcccaccca?cccacctatt?1740

aaccgcacct?acctacctac?ttacctacct?atccttcatc?gacctgcatc?aaaccgccgc?1800

ctcgacagac?agacgcccga?ctccatgaag?ctaacgtctc?cctcgcttcg?aggaacagct?1860

gtgttcatgc?ccgctccaac?aaggttctcc?atccgtcttt?catagaccct?gcccctttag?1920

ggggccaacc?gtctcgcttg?catctacgta?cacttcctcg?aaccgataac?cactgccagg?1980

aattatggcg?gaccgatcag?cacgccgtgc?gcgccctggc?gacgactctg?taggacagtt?2040

tgtcatcggg?gcggagatcg?gaaagggcag?cttcgcccaa?gtttacatgg?gcaagcacaa?2100

ggtcagcata?cattgacctc?ttcggattac?atgccatggc?aagcacccgt?tttagatcat?2160

attcgagacg?agctcgcgga?gcgaaatgca?ctagaacgat?gtgacaccgt?ccacgatggc?2220

tccccttcga?ctattggctg?tgccattcca?cagaaacatt?agtttgacga?ccgagtagct?2280

aacataacat?ttgcaccttg?acaggtatct?ggtgccgccg?ttgccatcaa?gtctgtcgag 2340

ctggcaagac?tcaataagaa?actcaaggag?aacctatacg?gggagataaa?tatcctgaag 2400

actctccgac?accctcatat?tgttgccctc?catgactgcg?tcgagtcggc?aacacacata 2460

aacctcatga?tggagtattg?cgagctaggg?gacctgtcgc?tattcattaa?gaagagggag 2520

aagctgtcga?cgaacccagc?aacacacgat?atggcccgca?agtaccccaa?cgtgccaaac 2580

tctggtctaa?acgaggtggt?cattcgacac?tttctcaaac?agctcagcag?cgctctgaaa 2640

ttccttagag?agagcaatct?tgtacaccga?gatgtgaagc?cgcagaacct?tctcctgctt 2700

ccctcgcccg?agttccggga?ggtaaacaaa?ttagcccgcc?ctatccttac?agcaagtcag 2760

gactccctcg?tcccagtcgc?cggccttgca?tcgcttccca?tgctgaagct?ggccgacttt 2820

ggctttgcga?gagtcctgcc?gtcaacgtca?ctggctgaga?ctctttgcgg?ctcaccattg 2880

tatatggccc?cagaaatcct?ccgttatgag?cggtacgatg?ccaaggcaga?cctctggtct 2940

gtcgggactg?tgttattcga?gatgatagtg?ggcaggccgc?ccttccgcgc?cagcaatcat 3000

gttgagctgc?tgaggaagat?cgaagccgct?gaggatgtga?tcaagttccc?ccgagagacc 3060

acgataagct?cagagatgaa?gggtcttaca?agggcacttc?tcaaacggaa?tcctgttgag 3120

cgcatcagct?tcgagaattt?ctttgctcac?ccagttatta?ttagttcgat?accgggcttg 3180

gttgaggatg?atatacccaa?gcctgaagct?agcgagcagc?gtagcagcag?caaggatacc 3240

agggcggcat?ccaaaagtga?tgacccaatc?gcttccccta?gaaagtattc?tttccgacgc 3300

cacccaacag?ataacgacca?aatcagggac?cagtttcgaa?gggtggaacc?gccgtcttct 3360

gctgcagaga?gcgccccttc?acggcaaaca?tcagctttta?gcggtattgc?cgcggaggct 3420

agaaaacaag?cagccgctga?agcatcagca?cgaaccggcc?agtcatcacg?caatgagcct 3480

ggagacaact?tggtccctcg?caggccgcaa?gcgcagcctt?ctacatcggc?tccgagcaag 3540

ccggggttgt?acgaagaacg?tcgccgtgga?atatcgaacg?catcgctgaa?tcgatccaac 3600

cgcgaatcat?catccccaac?tagtgcagct?ttggccaatg?attctgcaag?agcgccaccg 3660

cagcaaactt?ctaggagagt?gcaggcggag?gagagagaga?aggccgcgca?ggatgtagcc 3720

tttgagcggg?actacgtcgt?tgtagagaag?aagcatgtcg?aggtcaacgc?tttcgcagat 3780

gagatggctg?ccaatcctag?gctgggcggc?caaggcacgc?cactgtcgcc?caagtctggg 3840

cagattgtcc?gaagagcaac?ccagcaggga?aatcctactt?ctaccactgg?cgcaattcct 3900

gcagcgccat?cgcgcacaat?gcagatcgcg?caaggcactc?agagacacta?tcacgagcgc 3960

ggaacttctc?tctcagccag?tccaggctca?accgcttcct?tcattaccaa?ggctatccaa 4020

gatgcaagct?tacgactgct?cggaattaag?tacccggccg?gtctgacaaa?gggtgcatcg 4080

ccacccgagt?tatataaccc?ctaccctgcc?taccctaccc?catccccacc?ggttggcctc 4140

ataagcagcg?gaaagcagag?tactcccgta?gacgaggatg?cgcgggttgc?gcagctgatc 4200

gaggagcacg?ccactcgcag?tgacgtcgtt?tatggctttg?cagaggtcaa?gtacaaacag 4260

ctggttcctc?tcgcaccatc?ggcggaacat?gggctaggag?gaacaacgct?cgaagacatg 4320

cccaccggcg?aggaagatgt?tctcacggtt?gaggccatag?tctctctgtc?tgaagaggca 4380

ctggtgctat?acgtaaaagc?cttgactctc?cttgcgaagt?cgatggatat?tgcaagcctg 4440

tggtgggcta?ggaagaaccg?aggtgatcaa?gcaaacaacg?ctctttcgtc?gacgagggac 4500

tctgtgaaca?cgcagacact?atcattgaga?ataaattcgg?ccgttcagtg?ggtccgctca 4560

agattcaatg?aggtgctcga?aaaggccgaa?gtagtcaggc?tccggttgat?ggacgcacag 4620

aagcgacttc?cagatgatca?tccgagtcat?cctagcaatc?acccacaagg?atctgaatca 4680

gtaaacggcg?caagcgcaga?aggagtcttc?ttgacagtcg?gtgtgaccgc?ggagaagctc 4740

atgtacgacc?gcgctctgga?gatgagtcgc?actgctgcga?tcaacgagat?caccaatgag 4800

gatttggccg?ggtgtgagat?ctcttatgtc?acggcgattc?gtatgctaga?ggctgttctc 4860

gacaatgacg?aggatgcccc?gaagcgaaga?ttgtctgcca?acatggatga?cagcggtggt 4920

gacggagaag?atggtcatgc?ggagatcaat?gccgatgacc?aacaggcggt?ccagaagagt 4980

gagttttgct?gtgttgaaac?cttttttttt?tttcgctccc?atcaaatgtt?tgctgggtac 5040

tgacagacta?ttttcatcgc?ttagtgattc?acatgattag?gtctcgtctt?acctccgtac 5100

gcagcaaggt?ccgcatgata?tccaatgcct?caaaggccca?gcaacaatct?cagccacaga 5160

gcctcatcag?gcgtcgcagc?ggtgatgtga?cgccgcgaag?tgtcccttca?tacagctctt 5220

gatgcactta?gaaacactcg?ggctcaaccg?ggtattgttt?catctttttc?acgacttgac 5280

gataactttc?acattcacga?acttgctttt?attattcatc?atttttacca?gattgctgcc 5340

tcagggcgtt?ttacgtttcg?gagctgatac?agttcacgat?aagcacccag?cactagttgg 5400

actacggggc?gcaacttggt?gctgaggacg?agcggcgcat?gaagtgctat?tctacgatga 5460

tttttcactg?tcatactggg?gagttctttt?tttttttttt?tcctccctcg?tttattggtc 5520

aactgggtat?tgttttactt?atgtccttgc?cttttttttc?ttttacgtat?ggagcaggag 5580

tataatttta?tttcgcagac?atagggtctg?gacttctgtt?ttttcttttc?cactcctcgg 5640

ctatgcctga?agcgttcagt?tccagatttt?ggatcgttcg?aatactgtac?ttttttttat 5700

tatttttctt?ctcattcatt?tgctgcggcc?aatgtcatga?tgacactgga?gaagagacgt 5760

cgacgggcgg?ttgcgggcat?atatagctgt?tacaagaata?tattagtgag?gtggatgcgg 5820

taatcgcttg?tgccccaaga?tatgccgtac?aaatgtgttc?caatgtctgt?cttactgaag 5880

ctatagttgc?tgcatcgatt?gggattggct?gtcacctgtg?acataataaa?cgggttgtac 5940

attgatccat?tgtcatatat?ataacagttt?aaattaatca?atacagtagt?gcgttcagtg 6000

aacgaggagg?ctgtagagtg?attgtctgtc?caaacaagtc?aacacaatgt?aggattagga 6060

tacccagggt?aagataaggt?atcgaatatc?ctactgccat?gagatcgtac?atcgtcgata 6120

tagccagcag?cttgctaact?tagttgtagg?cataccaatt?gagcgctata?tctgcctcct 6180

atacccatac?tcactcctgc?tcctggg 6207

SEQ?ID?NO：2

atggcggacc?gatcagcacg?ccgtgcgcgc?cctggcgacg?actctgtagg?acagtttgtc 60

atcggggcgg?agatcggaaa?gggcagcttc?gcccaagttt?acatgggcaa?gcacaaggta 120

tctggtgccg?ccgttgccat?caagtctgtc?gagctggcaa?gactcaataa?gaaactcaag 180

gagaacctat?acggggagat?aaatatcctg?aagactctcc?gacaccctca?tattgttgcc 240

ctccatgact?gcgtcgagtc?ggcaacacac?ataaacctca?tgatggagta?ttgcgagcta 300

ggggacctgt?cgctattcat?taagaagagg?gagaagctgt?cgacgaaccc?agcaacacac 360

gatatggccc?gcaagtaccc?caacgtgcca?aactctggtc?taaacgaggt?ggtcattcga 420

cactttctca?aacagctcag?cagcgctctg?aaattcctta?gagagagcaa?tcttgtacac 480

cgagatgtga?agccgcagaa?ccttctcctg?cttccctcgc?ccgagttccg?ggaggtaaac 540

aaattagccc?gccctatcct?tacagcaagt?caggactccc?tcgtcccagt?cgccggcctt 600

gcatcgcttc?ccatgctgaa?gctggccgac?tttggctttg?cgagagtcct?gccgtcaacg 660

tcactggctg?agactctttg?cggctcacca?ttgtatatgg?ccccagaaat?cctccgttat 720

gagcggtacg?atgccaaggc?agacctctgg?tctgtcggga?ctgtgttatt?cgagatgata 780

gtgggcaggc?cgcccttccg?cgccagcaat?catgttgagc?tgctgaggaa?gatcgaagcc 840

gctgaggatg?tgatcaagtt?cccccgagag?accacgataa?gctcagagat?gaagggtctt 900

acaagggcac?ttctcaaacg?gaatcctgtt?gagcgcatca?gcttcgagaa?tttctttgct 960

cacccagtta?ttattagttc?gataccgggc?ttggttgagg?atgatatacc?caagcctgaa 1020

gctagcgagc?agcgtagcag?cagcaaggat?accagggcgg?catccaaaag?tgatgaccca 1080

atcgcttccc?ctagaaagta?ttctttccga?cgccacccaa?cagataacga?ccaaatcagg 1140

gaccagtttc?gaagggtgga?accgccgtct?tctgctgcag?agagcgcccc?ttcacggcaa 1200

acatcagctt?ttagcggtat?tgccgcggag?gctagaaaac?aagcagccgc?tgaagcatca 1260

gcacgaaccg?gccagtcatc?acgcaatgag?cctggagaca?acttggtccc?tcgcaggccg 1320

caagcgcagc?cttctacatc?ggctccgagc?aagccggggt?tgtacgaaga?acgtcgccgt 1380

ggaatatcga?acgcatcgct?gaatcgatcc?aaccgcgaat?catcatcccc?aactagtgca 1440

gctttggcca?atgattctgc?aagagcgcca?ccgcagcaaa?cttctaggag?agtgcaggcg 1500

gaggagagag?agaaggccgc?gcaggatgta?gcctttgagc?gggactacgt?cgttgtagag 1560

aagaagcatg?tcgaggtcaa?cgctttcgca?gatgagatgg?ctgccaatcc?taggctgggc 1620

ggccaaggca?cgccactgtc?gcccaagtct?gggcagattg?tccgaagagc?aacccagcag 1680

ggaaatccta?cttctaccac?tggcgcaatt?cctgcagcgc?catcgcgcac?aatgcagatc 1740

gcgcaaggca?ctcagagaca?ctatcacgag?cgcggaactt?ctctctcagc?cagtccaggc 1800

tcaaccgctt?ccttcattac?caaggctatc?caagatgcaa?gcttacgact?gctcggaatt 1860

aagtacccgg?ccggtctgac?aaagggtgca?tcgccacccg?agttatataa?cccctaccct 1920

gcctacccta?ccccatcccc?accggttggc?ctcataagca?gcggaaagca?gagtactccc 1980

gtagacgagg?atgcgcgggt?tgcgcagctg?atcgaggagc?acgccactcg?cagtgacgtc 2040

gtttatggct?ttgcagaggt?caagtacaaa?cagctggttc?ctctcgcacc?atcggcggaa 2100

catgggctag?gaggaacaac?gctcgaagac?atgcccaccg?gcgaggaaga?tgttctcacg 2160

gttgaggcca?tagtctctct?gtctgaagag?gcactggtgc?tatacgtaaa?agccttgact 2220

ctccttgcga?agtcgatgga?tattgcaagc?ctgtggtggg?ctaggaagaa?ccgaggtgat 2280

caagcaaaca?acgctctttc?gtcgacgagg?gactctgtga?acacgcagac?actatcattg 2340

agaataaatt?cggccgttca?gtgggtccgc?tcaagattca?atgaggtgct?cgaaaaggcc 2400

gaagtagtca?ggctccggtt?gatggacgca?cagaagcgac?ttccagatga?tcatccgagt 2460

catcctagca?atcacccaca?aggatctgaa?tcagtaaacg?gcgcaagcgc?agaaggagtc 2520

ttcttgacag?tcggtgtgac?cgcggagaag?ctcatgtacg?accgcgctct?ggagatgagt 2580

cgcactgctg?cgatcaacga?gatcaccaat?gaggatttgg?ccgggtgtga?gatctcttat 2640

gtcacggcga?ttcgtatgct?agaggctgtt?ctcgacaatg?acgaggatgc?cccgaagcga 2700

agattgtctg?ccaacatgga?tgacagcggt?ggtgacggag?aagatggtca?tgcggagatc 2760

aatgccgatg?accaacaggc?ggtccagaag?atgattcaca?tgattaggtc?tcgtcttacc 2820

tccgtacgca?gcaaggtccg?catgatatcc?aatgcctcaa?aggcccagca?acaatctcag 2880

ccacagagcc?tcatcaggcg?tcgcagcggt?gatgtgacgc?cgcgaagtgt?cccttcatac 2940

agctcttga 2949

SEQ?ID?NO：3

Met?Ala?Asp?Arg?Ser?Ala?Arg?Arg?Ala?Arg?Pro?Gly?Asp?Asp?Ser?Val?Gly?Gln?Phe?Val

Ile?Gly?Ala?Glu?Ile?Gly?Lys?Gly?Ser?Phe?Ala?Gln?Val?Tyr?Met?Gly?Lys?His?Lys?Val

Ser?Gly?Ala?Ala?Val?Ala?Ile?Lys?Ser?Val?Glu?Leu?Ala?Arg?Leu?Asn?Lys?Lys?Leu?Lys

Glu?Asn?Leu?Tyr?Gly?Glu?Ile?Asn?Ile?Leu?Lys?Thr?Leu?Arg?His?Pro?His?Ile?Val?Ala

Leu?His?Asp?Cys?Val?Glu?Ser?Ala?Thr?His?Ile?Asn?Leu?Met?Met?Glu?Tyr?Cys?Glu?Leu

Gly?Asp?Leu?Ser?Leu?Phe?Ile?Lys?Lys?Arg?Glu?Lys?Leu?Ser?Thr?Asn?Pro?Ala?Thr?His

Asp?Met?Ala?Arg?Lys?Tyr?Pro?Asn?Val?Pro?Asn?Ser?Gly?Leu?Asn?Glu?Val?Val?Ile?Arg

His?Phe?Leu?Lys?Gln?Leu?Ser?Ser?Ala?Leu?Lys?Phe?Leu?Arg?Glu?Ser?Asn?Leu?Val?His

Arg?Asp?Val?Lys?Pro?Gln?Asn?Leu?Leu?Leu?Leu?Pro?Ser?Pro?Glu?Phe?Arg?Glu?Val?Asn

Lys?Leu?Ala?Arg?Pro?Ile?Leu?Thr?Ala?Ser?Gln?Asp?Ser?Leu?Val?Pro?Val?Ala?Gly?Leu

Ala?Ser?Leu?Pro?Met?Leu?Lys?Leu?Ala?Asp?Phe?Gly?Phe?Ala?Arg?Val?Leu?Pro?Ser?Thr

Ser?Leu?Ala?Glu?Thr?Leu?Cys?Gly?Ser?Pro?Leu?Tyr?Met?Ala?Pro?Glu?Ile?Leu?Arg?Tyr

Glu?Arg?Tyr?Asp?Ala?Lys?Ala?Asp?Leu?Trp?Ser?Val?Gly?Thr?Val?Leu?Phe?Glu?Met?Ile

Val?Gly?Arg?Pro?Pro?Phe?Arg?Ala?Ser?Asn?His?Val?Glu?Leu?Leu?Arg?Lys?Ile?Glu?Ala

Ala?Glu?Asp?Val?Ile?Lys?Phe?Pro?Arg?Glu?Thr?Thr?Ile?Ser?Ser?Glu?Met?Lys?Gly?Leu

Thr?Arg?Ala?Leu?Leu?Lys?Arg?Asn?Pro?Val?Glu?Arg?Ile?Ser?Phe?Glu?Asn?Phe?Phe?Ala

His?Pro?Val?Ile?Ile?Ser?Ser?Ile?Pro?Gly?Leu?Val?Glu?Asp?Asp?Ile?Pro?Lys?Pro?Glu

Ala?Ser?Glu?Gln?Arg?Ser?Ser?Ser?Lys?Asp?Thr?Arg?Ala?Ala?Ser?Lys?Ser?Asp?Asp?Pro

Ile?Ala?Ser?Pro?Arg?Lys?Tyr?Ser?Phe?Arg?Arg?His?Pro?Thr?Asp?Asn?Asp?Gln?Ile?Arg

Asp?Gln?Phe?Arg?Arg?Val?Glu?Pro?Pro?Ser?Ser?Ala?Ala?Glu?Ser?Ala?Pro?Ser?Arg?Gln

Thr?Ser?Ala?Phe?Ser?Gly?Ile?Ala?Ala?Glu?Ala?Arg?Lys?Gln?Ala?Ala?Ala?Glu?Ala?Ser

Ala?Arg?Thr?Gly?Gln?Ser?Ser?Arg?Asn?Glu?Pro?Gly?Asp?Asn?Leu?Val?Pro?Arg?Arg?Pro

Gln?Ala?Gln?Pro?Ser?Thr?Ser?Ala?Pro?Ser?Lys?Pro?Gly?Leu?Tyr?Glu?Glu?Arg?Arg?Arg

Gly?Ile?Ser?Asn?Ala?Ser?Leu?Asn?Arg?Ser?Asn?Arg?Glu?Ser?Ser?Ser?Pro?Thr?Ser?Ala

Ala?Leu?Ala?Asn?Asp?Ser?Ala?Arg?Ala?Pro?Pro?Gln?Gln?Thr?Ser?Arg?Arg?Val?Gln?Ala

Glu?Glu?Arg?Glu?Lys?Ala?Ala?Gln?Asp?Val?Ala?Phe?Glu?Arg?Asp?Tyr?Val?Val?Val?Glu

Lys?Lys?His?Val?Glu?Val?Asn?Ala?Phe?Ala?Asp?Glu?Met?Ala?Ala?Asn?Pro?Arg?Leu?Gly

Gly?Gln?Gly?Thr?Pro?Leu?Ser?Pro?Lys?Ser?Gly?Gln?Ile?Val?Arg?Arg?Ala?Thr?Gln?Gln

Gly?Asn?Pro?Thr?Ser?Thr?Thr?Gly?Ala?Ile?Pro?Ala?Ala?Pro?Ser?Arg?Thr?Met?Gln?Ile

Ala?Gln?Gly?Thr?Gln?Arg?His?Tyr?His?Glu?Arg?Gly?Thr?Ser?Leu?Ser?Ala?Ser?Pro?Gly

Ser?Thr?Ala?Ser?Phe?Ile?Thr?Lys?Ala?Ile?Gln?Asp?Ala?Ser?Leu?Arg?Leu?Leu?Gly?Ile

Lys?Tyr?Pro?Ala?Gly?Leu?Thr?Lys?Gly?Ala?Ser?Pro?Pro?Glu?Leu?Tyr?Asn?Pro?Tyr?Pro

Ala?Tyr?Pro?Thr?Pro?Ser?Pro?Pro?Val?Gly?Leu?Ile?Ser?Ser?Gly?Lys?Gln?Ser?Thr?Pro

Val?Asp?Glu?Asp?Ala?Arg?Val?Ala?Gln?Leu?Ile?Glu?Glu?His?Ala?Thr?Arg?Ser?Asp?Val

Val?Tyr?Gly?Phe?Ala?Glu?Val?Lys?Tyr?Lys?Gln?Leu?Val?Pro?Leu?Ala?Pro?Ser?Ala?Glu

His?Gly?Leu?Gly?Gly?Thr?Thr?Leu?Glu?Asp?Met?Pro?Thr?Gly?Glu?Glu?Asp?Val?Leu?Thr

Val?Glu?Ala?Ile?Val?Ser?Leu?Ser?Glu?Glu?Ala?Leu?Val?Leu?Tyr?Val?Lys?Ala?Leu?Thr

Leu?Leu?Ala?Lys?Ser?Met?Asp?Ile?Ala?Ser?Leu?Trp?Trp?Ala?Arg?Lys?Asn?Arg?Gly?Asp

Gln?Ala?Asn?Asn?Ala?Leu?Ser?Ser?Thr?Arg?Asp?Ser?Val?Asn?Thr?Gln?Thr?Leu?Ser?Leu

Arg?Ile?Asn?Ser?Ala?Val?Gln?Trp?Val?Arg?Ser?Arg?Phe?Asn?Glu?Val?Leu?Glu?Lys?Ala

Glu?Val?Val?Arg?Leu?Arg?Leu?Met?Asp?Ala?Gln?Lys?Arg?Leu?Pro?Asp?Asp?His?Pro?Ser

His?Pro?Ser?Asn?His?Pro?Gln?Gly?Ser?Glu?Ser?Val?Asn?Gly?Ala?Ser?Ala?Glu?Gly?Val

Phe?Leu?Thr?Val?Gly?Val?Thr?Ala?Glu?Lys?Leu?Met?Tyr?Asp?Arg?Ala?Leu?Glu?Met?Ser

Arg?Thr?Ala?Ala?Ile?Asn?Glu?Ile?Thr?Asn?Glu?Asp?Leu?Ala?Gly?Cys?Glu?Ile?Ser?Tyr

Val?Thr?Ala?Ile?Arg?Met?Leu?Glu?Ala?Val?Leu?Asp?Asn?Asp?Glu?Asp?Ala?Pro?Lys?Arg

Arg?Leu?Ser?Ala?Asn?Met?Asp?Asp?Ser?Gly?Gly?Asp?Gly?Glu?Asp?Gly?His?Ala?Glu?Ile

Asn?Ala?Asp?Asp?Gln?Gln?Ala?Val?Gln?Lys?Met?Ile?His?Met?Ile?Arg?Ser?Arg?Leu?Thr

Ser?Val?Arg?Ser?Lys?Val?Arg?Met?Ile?Ser?Asn?Ala?Ser?Lys?Ala?Gln?Gln?Gln?Ser?Gln

Pro?Gln?Ser?Leu?Ile?Arg?Arg?Arg?Ser?Gly?Asp?Val?Thr?Pro?Arg?Ser?Val?Pro?Ser?Tyr

Ser?Ser

SEQ?ID?NO：4

aagaagatgg?aggggacgct?aaatggtacc?gatctggcta?aagcctgcca?agcagccacc 60

aactcgtgac?agaggaacag?aatggcatct?gtgcgaagtc?tcttggttgc?ttctcaacga 120

atgcagtcgt?tcatcaggcg?ttctatttgc?tgcctcacat?aggcgagttt?gggtaggcag 180

cttaccttat?ctgtcgtgtt?gtcatttcgc?aaggaaagtc?gcaacaggtt?gtcatggatc 240

gtctaatcaa?ttacgtctat?cagttccaag?ctatcatcat?ctcaacctga?ctaagtgtgt 300

agtacccatc?tatgtatctt?tgggcatggc?atgcctaccg?cgtatcacac?cctcgaaatc 360

cacagtgcca?caattatcat?ccccaactat?gcagggcact?gtcgtttctc?atttacccat 420

cttatctgct?ctctgcgtcg?taaatctccg?tctttattgc?ctagtgcctt?tacacttctt 480

ttattagaca?ccgcgttggg?ggttctggaa?taacttactc?cgtacaatag?atagctttct 540

ttttgttctt?tctttctttc?tttttttttt?tttttttaaa?aaaaaaagat?aggaaaaaga 600

aagaaagaaa?gaaaagactg?cacttaatat?ctgaaatctc?agttcagtcc?aaggcaaggt 660

ctctaaaggt?acctacctag?ataggtacct?accaggtacg?taccttacgt?acctcactga 720

ctccgccaaa?ccccacactg?gtgtcgtcaa?cccaagttcc?gatcacgcac?cagtctttgc 780

ccattgctat?ggagctcacc?agacctgaca?cgccggggac?agtgcaggcg?tcacggttga 840

cgccaaatta?ggcatcccgg?acatgaagtt?tatgttacgg?tgttgtaaat?aattattaca 900

ctgacacgaa?tgcagccttt?tgacagaagt?caaccttcct?aggccacaaa?actcttatta 960

ctctgcttgc?tcggtcctag?atctggacgt?acagtaggta?attggtaggt?acctagttag 1020

gtaggtaggt?tactgtattg?taatggaggt?gaggtgatga?ccgtacagta?ccggtggtga 1080

ttgtgcagcc?agttcagtgg?catccgcccg?cccagtgccg?ttaaaaaaaa?aaaaaaccac 1140

ccgctcatta?gaccgcccac?ccccagctgc?cacctgccca?cgtcttacac?aaactccgaa 1200

aaacgaacga?cttaccctgt?ttgttaccac?tttcaattgc?cttatcttct?catcaaagcc 1260

gtcacgagtt?ggggatcaaa?ctcgaaaaaa?atcgaggcct?tcaaagacac?gtcttgaaag 1320

aggcttggcc?catttgcgaa?tagaaaaagg?tcggtatgtc?ttgtggtgac?gtcgttttcg 1380

ctttcaacct?gtcgccccct?taaactattg?tttacttctc?ggtcgcaggc?tatgcagcgg 1440

agcgctaggc?gcgtggaacc?ccctgccaca?agtcacttgc?gctccacatc?caatccccac 1500

acatgatcca?gaacctcgtc?gtgaatcctg?ggcctatcgg?agccgtgcca?gagaccaggc 1560

ggcggaccag?ttcctaccat?tcgtgacctt?gcctgcctgc?ctccccggcc?tagcgatcca 1620

ggcaccttgt?tgactgcttt?gaactgtgct?tgctcatcat?cacccaactt?ttctcttacc 1680

tatctttttt?ctgtttttcc?ctcccgagtc?ttttcatccc?atcccaccca?cccacctatt 1740

aaccgcacct?acctacctac?ttacctacct?atccttcatc?gacctgcatc?aaaccgccgc 1800

ctcgacagac?agacgcccga?ctccatgaag?ctaacgtctc?cctcgcttcg?aggaacagct 1860

gtgttcatgc?ccgctccaac?aaggttctcc?atccgtcttt?catagaccct?gcccctttag 1920

ggggccaacc?gtctcgcttg?catctacgta?cacttcctcg?aaccgataac?cactgccagg 1980

aatt 1984

SEQ?ID?NO：5

Met?Ala?Gly?Pro?Gln?Glu?Ser?Ser?Thr?Ser?Ser?Gly?Ser?Arg?Lys?Ser?Gly?Ser?Arg?Ala

Val?Gly?Gln?Phe?Asn?Ile?Gly?Ser?Glu?Ile?Gly?Lys?Gly?Ser?Phe?Ala?Gln?Val?Tyr?Leu

Gly?Trp?His?Lys?Glu?Thr?Lys?Ala?Ala?Val?Ala?Ile?Lys?Ser?Val?Glu?Leu?Glu?Arg?Leu

Asn?Lys?Lys?Leu?Arg?Glu?Asn?Leu?Tyr?Ser?Glu?Ile?Gln?Ile?Leu?Lys?Thr?Leu?Arg?His

Pro?His?Ile?Val?Ala?Leu?His?Asp?Cys?Ile?Glu?Ser?Thr?Ser?His?Ile?Asn?Leu?Ile?Met

Glu?Tyr?Cys?Glu?Leu?Gly?Asp?Leu?Ser?Leu?Phe?Ile?Lys?Lys?Arg?Glu?Lys?Leu?Ala?Thr

His?Pro?Ala?Thr?His?Asp?Met?Ala?Arg?Lys?Tyr?Pro?Ser?Met?Pro?Asn?Ser?Gly?Leu?His

Glu?Val?Val?Ile?Arg?His?Phe?Leu?Lys?Gln?Leu?Thr?Ser?Ala?Leu?Glu?Phe?Leu?Arg?Ser

Lys?Asn?Tyr?Val?His?Arg?Asp?Val?Lys?Pro?Gln?Asn?Leu?Leu?Leu?Leu?Pro?Ser?Gln?Pro

Phe?Arg?Asp?Gln?Arg?Ser?Arg?Pro?Val?Met?Gln?Ala?Ser?Gln?Asp?Ser?Leu?Ile?Pro?Ile

Ser?Gly?Leu?Ala?Ser?Leu?Pro?Met?Leu?Lys?Leu?Ala?Asp?Phe?Gly?Phe?Ala?Arg?Val?Leu

Pro?Ser?Thr?Ser?Leu?Ala?Asp?Thr?Leu?Cys?Gly?Ser?Pro?Leu?Tyr?Met?Ala?Pro?Glu?Ile

Leu?Arg?Tyr?Glu?Arg?Tyr?Asp?Ala?Lys?Ala?Asp?Leu?Trp?Ser?Val?Gly?Thr?Val?Leu?Tyr

Glu?Met?Ser?Thr?Gly?Arg?Pro?Pro?Phe?Arg?Ala?Arg?Asn?His?Val?Glu?Leu?Leu?Arg?Lys

Ile?Glu?Ala?Ala?Glu?Asp?Val?Ile?Lys?Phe?Pro?Arg?Glu?Val?Ser?Ile?Thr?Pro?Glu?Leu

Lys?Ala?Leu?Ile?Arg?Ser?Leu?Leu?Lys?Arg?Ser?Pro?Val?Glu?Arg?Leu?Ser?Phe?Glu?Asn

Phe?Phe?Thr?His?Gln?Val?Val?Thr?Ser?Glu?Ile?Pro?Gly?Leu?Val?Glu?Asp?Asp?Ile?Pro

Lys?Ser?Leu?Arg?Gln?Glu?Ser?Arg?Asp?Pro?Arg?Ser?Ala?Phe?Gln?Ser?Gly?Ser?Pro?Ser

Leu?Ser?Ser?Arg?Ser?Pro?Arg?Gln?Thr?Gly?His?Gln?Ser?Pro?Thr?Glu?Ala?Leu?Val?Ser

Arg?Ser?Pro?Arg?Asp?Gln?Gln?Pro?Arg?Ser?Pro?Gln?Val?Gly?Ser?Pro?Gly?Gly?Ser?Arg

Tyr?Ala?Arg?Arg?Ser?Asn?Glu?Ser?Gln?Arg?Thr?Thr?Gly?Asn?Ser?Pro?Arg?Glu?Gly?Gly

Glu?Gly?Leu?Gly?Ile?Arg?Arg?Pro?Val?Ala?Gln?His?Ala?Met?Thr?Ala?Pro?Val?Gln?Gln

Val?Ala?Tyr?Asp?Ser?Val?Thr?Gly?Arg?Asn?Arg?Ala?Ser?Pro?Pro?Thr?Ser?Leu?Leu?Asp

Gln?Val?Arg?Arg?Asn?Arg?Ala?Leu?Ser?Asn?Pro?Pro?Ile?Thr?Glu?Glu?Glu?Arg?Ala?Ala

Gln?Asp?Val?Ala?Leu?Glu?Arg?Glu?Tyr?Val?Val?Val?Glu?Arg?Arg?His?Val?Glu?Val?Asn

Ala?Leu?Ala?Asp?Glu?Leu?Ala?Ala?Asn?Glu?Lys?Leu?Gly?Asp?Ala?Ser?Gln?Arg?Ser?Gly

Pro?Ile?Thr?Arg?Arg?Tyr?Thr?Gln?Gln?Gly?Ala?Pro?Thr?Ser?Thr?Thr?Gly?Ala?Ile?Ser

Thr?Pro?Tyr?Ser?Arg?Asn?Ala?Leu?Ala?Thr?Gln?Pro?Arg?His?Asp?Arg?Lys?Ser?Ser?Tyr

Glu?Lys?Ser?Leu?Ser?Ala?Ser?Pro?Gly?Ser?Ala?Ser?Ser?Ala?Ile?Ser?Lys?Ala?Ile?Gln

Asp?Ala?Ser?Leu?Arg?Leu?Phe?Gly?Phe?Lys?Val?Pro?Pro?Leu?Arg?Ala?Ser?Pro?Lys?Gly

Pro?Ser?Pro?Pro?Leu?Tyr?Gln?Ala?Phe?Pro?Thr?Tyr?Pro?Thr?Pro?Gln?Ala?Pro?Val?Gly

Leu?Leu?Gly?Asp?Gly?Arg?Asn?Val?Gln?Gly?Thr?Asp?Glu?Asp?Gly?Lys?Ala?Ala?Gln?Thr

Ile?Glu?Glu?Leu?Ala?Thr?Arg?Ser?Asp?Cys?Val?Tyr?Gly?Phe?Ala?Glu?Val?Lys?Tyr?Lys

Gln?Leu?Val?Pro?Leu?Ala?Pro?Ser?Ala?Asp?His?Ile?Leu?Gly?Gly?Leu?Glu?Pro?Glu?Gln

Leu?Val?Asn?Glu?Glu?Asp?Gly?Leu?Thr?Val?Glu?Ala?Ile?Val?Ala?Leu?Ser?Glu?Glu?Ala

Leu?Val?Leu?Tyr?Val?Lys?Ser?Leu?Thr?Leu?Leu?Ala?Arg?Ala?Met?Asp?Ile?Ala?Ser?Leu

Trp?Trp?Ser?Lys?Lys?Ser?Arg?Gly?Asp?Thr?Gly?Thr?Gly?Leu?Ser?Ala?Ala?Ala?Ala?Gln

Thr?Val?Val?Gln?Arg?Ile?Asn?Ala?Val?Val?Gln?Trp?Val?Arg?Gln?Arg?Phe?Asn?Glu?Val

Leu?Glu?Lys?Ser?Glu?Ile?Val?Arg?Leu?Lys?Leu?Thr?Glu?Ala?Gln?Lys?Gln?Leu?Pro?Asp

Asp?His?Pro?Ser?His?Pro?Ser?Asn?His?Gly?Thr?Glu?Ser?Ile?Ala?Ser?Ser?Ala?Gly?Ser

Pro?Thr?Lys?Gln?Val?Tyr?Leu?Thr?Pro?Gly?Ile?Ser?Ala?Glu?Lys?Leu?Met?Tyr?Asp?Arg

Ala?Leu?Glu?Met?Ser?Arg?Ala?Ala?Ala?Ile?Asp?Glu?Val?Thr?Asn?Glu?Asn?Leu?Ser?Gly

Cys?Glu?Ile?Ser?Tyr?Ile?Thr?Ala?Ile?Arg?Met?Leu?Glu?Ala?Val?Leu?Asp?Asn?Asp?Glu

Gly?Ser?Gly?Ser?Glu?Thr?Arg?Arg?Leu?Ser?Thr?Gly?Lys?Glu?Ala?Glu?Arg?Glu?Ala?Val

Lys?Glu?Val?Ser?Gly?Gly?Glu?Leu?Asp?Ser?Asp?Glu?Glu?Ala?His?Val?Arg?Lys?Arg?Arg

Leu?Ala?Ala?Val?Arg?Lys?Lys?Gln?Gln?Met?Ile?Ala?Glu?Ala?Asn?Ser?Lys?Thr?Asn?Leu

Val?Tyr?Gln?Gln?Ala?Val?Arg?Arg?Arg?Ser?Gly?Asp?Met?Thr?Pro?Arg?Ser?Val?Pro?Ser

His?Ala?Ser?Ser

SEQ?ID?NO：6

Met?Ala?Ser?Thr?Thr?Thr?Ser?Thr?Ser?Ser?Leu?Ser?Ser?Arg?Arg?Gln?Lys?Thr?Gly?Val

Gly?Ser?Phe?Thr?Ile?Asn?Glu?Gln?Ile?Gly?Lys?Gly?Ser?Phe?Ala?Thr?Val?Tyr?Arg?Gly

Thr?His?Met?Pro?Ser?Gly?Asn?Leu?Val?Ala?Ile?Lys?Ser?Val?Asn?Leu?Ser?Arg?Leu?Asn

Lys?Lys?Leu?Lys?Asp?Asn?Leu?Tyr?Val?Glu?Ile?Glu?Ile?Leu?Lys?Ser?Leu?Tyr?His?Pro

His?Ile?Val?Ala?Leu?Ile?Asp?Cys?Arg?Glu?Ser?Ala?Ser?His?Ile?His?Leu?Met?Met?Glu

Tyr?Cys?Glu?Leu?Gly?Asp?Leu?Ser?Tyr?Phe?Ile?Lys?Lys?Arg?Asp?Arg?Leu?Ala?Asp?Asn

Pro?Thr?Leu?Tyr?Asp?Met?Val?Gln?Lys?Tyr?Pro?Met?Pro?Val?Glu?Gly?Gly?Leu?Asn?Gln

Val?Val?Val?Arg?His?Phe?Phe?Lys?Gln?Leu?Ser?Ser?Ala?Met?Glu?Phe?Leu?Arg?Glu?Arg

Asp?Phe?Val?His?Arg?Asp?Val?Lys?Pro?Gln?Asn?Leu?Leu?Leu?Ile?Pro?Ser?Pro?Glu?Trp

Ile?Ala?Lys?Arg?Ala?Lys?Gly?Gly?Pro?Glu?Ala?Met?Lys?Ala?Ser?Lys?Glu?Ser?Val?Val

Ala?Met?Val?Gly?Ile?Asn?Ser?Leu?Pro?Met?Leu?Lys?Leu?Ala?Asp?Phe?Gly?Phe?Ala?Arg

Ser?Leu?Pro?Ser?Thr?Ser?Leu?Ala?Glu?Thr?Leu?Cys?Gly?Ser?Pro?Leu?Tyr?Met?Ala?Pro

Glu?Ile?Leu?Arg?Tyr?Glu?Lys?Tyr?Asp?Ala?Arg?Ala?Asp?Leu?Trp?Ser?Ile?Gly?Thr?Val

Leu?Tyr?Glu?Met?Met?Thr?Gly?Arg?Pro?Pro?Phe?Lys?Ala?Ile?Asn?His?Val?Gln?Leu?Leu

Gln?Lys?Ile?Glu?Lys?Asn?Gln?Asp?Glu?Ile?Arg?Phe?Pro?Ser?Arg?Gly?Ile?Tyr?Ser?Arg

Asp?Leu?Lys?Asp?Ile?Val?Arg?Arg?Leu?Leu?Lys?Lys?Lys?Pro?Glu?Asp?Arg?Ile?Thr?Phe

Pro?Glu?Tyr?Phe?Ala?His?Pro?Val?Val?Thr?Glu?Pro?Ile?Pro?Gly?Leu?Val?Gly?Asp?Asp

Arg?Pro?Lys?Glu?Lys?Ser?Pro?Glu?Thr?Ser?Ile?Val?Arg?Gln?Pro?Ser?Leu?Arg?Asp?Arg

Gln?Arg?Glu?Ser?Pro?Thr?Val?Lys?His?Ile?Asp?Thr?Ala?Tyr?Glu?Ser?Leu?Ile?Thr?Arg

Asp?Ile?Gly?Glu?Gln?Ser?Pro?Arg?Thr?Pro?Asn?Ile?Glu?Ser?Asn?Gln?Pro?Phe?Gly?Thr

Pro?Gly?Arg?Ser?Ser?Gly?Arg?Pro?Asp?Ser?Arg?Asp?Arg?Pro?Ser?Pro?Val?Ser?Ala?Ala

Thr?Ala?Pro?Asn?Val?Asp?Thr?Leu?Pro?Arg?Gln?Arg?Asp?Arg?Lys?Asp?Arg?Thr?Glu?Pro

Asn?Tyr?Ala?Pro?Ile?Val?Arg?Thr?Gly?Ser?Thr?Lys?Gln?Arg?Tyr?Asp?Glu?Gln?Ala?Asn

Leu?Gln?Pro?Lys?Asn?Glu?Val?Gln?Ser?Ser?Asn?Ser?Ile?Thr?Glu?Ala?Glu?Gln?Asp?Val

Arg?Asp?Ala?Arg?Glu?Tyr?Val?Leu?Val?Glu?Lys?Lys?Ala?Val?Glu?Val?Asn?Ala?Phe?Ala

Asp?Glu?Met?Ala?Ala?Asn?Pro?Arg?Leu?Gly?Arg?Ala?Asn?Ser?Ala?Pro?Lys?Gln?Leu?Pro

Arg?Arg?His?Thr?Ser?Met?Gly?Glu?Pro?Asn?Ser?Thr?Thr?Gly?Ala?Val?Ala?Val?Pro?Pro

Ser?Arg?Ile?Val?Gln?Arg?Ala?Ser?Gly?Arg?Ala?Gln?Pro?Asp?Thr?Ser?Ser?Ala?Arg?Asn

Ser?Tyr?Gly?Ssr?Tyr?Gly?Lys?Thr?Gly?Ser?Ser?Pro?Ser?Thr?Ala?Ser?Ala?Ile?Ala?Lys

Ala?Leu?Gln?Gly?Ala?Ser?Val?Arg?Val?Phe?Gly?Val?Ser?Trp?Ser?Pro?Thr?Leu?Ile?Gly

Lys?Gly?Pro?Ser?Pro?Pro?Gln?Leu?Tyr?Asn?Pro?Tyr?Pro?Ala?Tyr?Pro?Thr?Pro?Asn?Ala

Gly?Leu?Ile?Gly?Asp?Gly?Arg?Pro?Ile?Asp?Glu?Asp?Gln?Arg?Val?Val?Asn?Ile?Ile?Glu

Asp?Ser?Ala?Thr?Arg?Ser?Asp?Val?Val?Tyr?Gly?Phe?Ala?Glu?Val?Lys?Tyr?Arg?Gln?Leu

Ile?Pro?Leu?Ala?Pro?Ser?Met?Asn?His?Gly?Leu?Gly?Gly?Pro?Asn?Pro?Glu?Arg?Thr?Gly

Asp?Ala?Met?Asp?Glu?Asp?Asp?Gly?Leu?Thr?Val?Glu?Ala?Ile?Val?Asn?Leu?Ser?Glu?Glu

Ala?Leu?Val?Leu?Tyr?Val?Lys?Ser?Leu?Ser?Leu?Leu?Ser?Lys?Ser?Met?Asp?Ile?Ala?Gly

Ala?Trp?Trp?Ser?Arg?Lys?Gln?Arg?Gly?Gly?Ile?Val?Ser?Gly?Gly?His?Thr?Pro?Gly?Ser

Asp?Ser?Ser?Ser?Ala?Ala?Gln?Ala?Gly?Asn?Arg?Ile?Asn?Gly?Ala?Val?Gln?Trp?Val?Arg

Thr?Arg?Phe?Asn?Glu?Val?Leu?Glu?Lys?Ala?Glu?Leu?Val?Arg?Leu?Lys?Leu?Val?Glu?Ala

Gln?Lys?Arg?Leu?Pro?Glu?Asp?His?Pro?Gly?His?Pro?Asn?Asn?Arg?Ser?Thr?Ala?Ser?Arg

Leu?Val?Gly?Gly?Ser?Ser?Thr?Thr?Asp?Gly?Val?Val?Leu?Ser?Ser?Gly?Ile?Thr?Ala?Glu

Lys?Leu?Met?Tyr?Asp?Arg?Ala?Leu?Glu?Met?Ser?Arg?Thr?Ala?Ala?Ile?Asn?Glu?Leu?Ala

Asn?Glu?Asp?Leu?Pro?Gly?Cys?Glu?Ile?Ser?Tyr?Thr?Thr?Ala?Ile?Arg?Met?Leu?Glu?Ala

Val?Leu?Glu?Asn?Asp?Glu?Glu?Leu?Ile?Pro?Arg?Lys?Arg?Ser?Ser?Ser?Leu?Arg?Glu?Asp

Lys?Glu?Lys?Ser?Glu?Gly?Gly?Glu?Val?Asn?Gly?Ile?Asn?Phe?Gly?Asp?Arg?Lys?Asp?Val

Leu?Lys?Val?Leu?Gln?Met?Ile?Arg?Thr?Arg?Leu?Gln?Val?Leu?Lys?Lys?Lys?Met?Thr?Ala

Ile?Ala?Lys?His?Gln?Ser?Met?Pro?Pro?Pro?Ser?Ser?Ser?Pro?Arg?Arg?Ser?Tyr?Ser?Gly

Gly?Thr?Thr?Pro?Thr?Ile?Asn?Asn?Thr?Pro?Pro?Lys

SEQ?ID?NO：7

Met?Ala?Ala?Leu?Gln?Ser?Pro?Lys?Thr?Ser?Ser?Arg?Arg?Ser?Lys?Gly?Asp?Gly?Asp?Gly

Asp?Thr?Arg?Asp?Val?Ile?Leu?Gly?Lys?Tyr?Thr?Lys?Ile?Glu?Glu?Ile?Gly?Arg?Gly?Ser

Phe?Ala?Thr?Val?Tyr?Gln?Gly?Ile?His?Asn?Lys?Tyr?Arg?Ser?Cys?Val?Ala?Ile?Lys?Ala

Val?Asn?Ile?Ser?Ser?Leu?Asn?Pro?Lys?Leu?Lys?Asp?Asn?Leu?Lys?Leu?Glu?Ile?Glu?Ile

Leu?Lys?Gly?Leu?Gln?His?Pro?His?Ile?Val?Ala?Leu?Ile?Asp?Cys?Asp?Glu?Ser?Thr?Ser

Cys?Ile?His?Leu?Val?Met?Glu?Tyr?Cys?Ala?Leu?Gly?Asp?Leu?Ser?Leu?Phe?Ile?Arg?Lys

Arg?Asp?Thr?Leu?Ser?Lys?His?Glu?Leu?Thr?Arg?Asp?Met?Ile?Ala?Lys?Tyr?Pro?Asn?Pro

Pro?Ala?Gly?Gly?Leu?Asn?Glu?Val?Ile?Val?Arg?His?Phe?Leu?Lys?Gln?Leu?Ala?Ser?Ala

Leu?Gln?Phe?Leu?Arg?Thr?Lys?Asp?Leu?Ile?His?Arg?Asp?Leu?Lys?Pro?Gln?Asn?Leu?Leu

Leu?Asn?Pro?Pro?Pro?Ser?Thr?Tyr?Ala?Lys?Gly?Leu?Leu?Arg?Ile?Val?Pro?Tyr?Lys?Thr

Arg?Glu?Asp?Ser?Phe?Thr?Pro?Leu?Val?Gly?Val?Glu?Ser?Leu?Pro?Met?Leu?Lys?Ile?Ala

Asp?Phe?Gly?Phe?Ala?Arg?Ser?Leu?Pro?Ala?Thr?Ser?Leu?Ala?Glu?Thr?Leu?Cys?Gly?Ser

Pro?Leu?Tyr?Met?Ala?Pro?Glu?Ile?Leu?Arg?Tyr?Glu?Lys?Tyr?Asp?Ala?Lys?Ala?Asp?Leu

Trp?Ser?Val?Gly?Thr?Val?Leu?Phe?Glu?Met?Val?Val?Gly?Lys?Ser?Pro?Phe?Arg?Ala?Gly

Asn?His?Val?Asp?Leu?Leu?Arg?Lys?Ile?Glu?Gln?Gly?Glu?Asp?Asn?Ile?Arg?Phe?Pro?Ile

Gln?Thr?Glu?Ala?Ser?Pro?Pro?Leu?Lys?Lys?Leu?Ile?Arg?Ser?Leu?Leu?Lys?Arg?Asn?pro

Val?Glu?Arg?Leu?Ser?Phe?Lys?Asp?Phe?Phe?Glu?Ser?Ser?Val?Val?Lys?Gly?Asn?Ile?Pro

Gly?Leu?Val?Asp?Glu?Asp?Leu?Gln?Ala?Gln?Arg?Glu?Arg?Asp?Ser?Arg?Leu?Ala?Ala?His

Ala?Ala?Arg?Arg?Asp?Ser?Leu?Pro?Ser?Arg?Thr?Ser?Asp?Gly?Lys?Leu?Glu?Asp?Arg?Pro

Pro?Ser?Ser?Ala?Ser?Ser?Pro?Arg?Pro?Pro?Pro?Ser?Asn?Phe?Thr?Arg?Pro?Ala?Thr?Ser

Pro?Ala?Val?Arg?His?Val?Gly?Ser?Arg?Glu?Asn?Pro?Asp?Leu?Gln?Arg?Val?Ala?Val?Lys

Ala?Arg?Lys?Ala?Ser?Val?Ala?Ser?Val?Thr?Gly?Ser?Pro?Ser?Lys?Glu?Glu?Leu?Arg?Asp

His?Asn?Ala?Lys?Gly?Pro?Val?Thr?Glu?Gln?Pro?Gly?Pro?Thr?Ser?Pro?Ala?Lys?Glu?Arg

Ala?Thr?Thr?Arg?Lys?Asn?Ser?Ser?Asp?Gln?Arg?Ile?Asp?Asp?Ile?Leu?Asp?Ser?Glu?Arg

Glu?Arg?Ala?Ala?Gln?Asp?Val?Ala?Phe?Glu?Arg?Asp?Tyr?Val?Val?Val?Glu?Lys?Arg?Ala

Val?Glu?Val?Asn?Ala?Phe?Ala?Asp?Glu?Leu?Ala?Ala?Ser?Pro?Arg?Phe?Gln?Gln?Gln?Gln

Gln?Gln?Gln?Leu?Ile?Pro?Lys?Gln?Ala?Gly?Ala?Ile?Val?Arg?Arg?Ala?Thr?Thr?Thr?Ala

Thr?Pro?Gln?Leu?Ser?Ser?Ser?Pro?Arg?Asp?Ala?Met?Ser?Arg?Ala?Val?Ala?Thr?Ala?Tyr

Asn?Arg?Pro?Arg?Thr?Gly?Ser?Thr?His?Asn?Arg?His?Asn?Ser?Tyr?Asp?Arg?Arg?Tyr?Gly

Pro?Ser?Pro?Thr?Ser?Ala?Thr?Ser?AlaIle?Ser?Lys?Ala?Leu?Asn?Lys?Ala?Ser?Gly?Arg

Leu?Phe?Gly?Val?Ser?Phe?Ser?Pro?Pro?Leu?Ala?Leu?Thr?Lys?Ala?Gly?Arg?Ser?Pro?Pro

Ile?Gly?Tyr?Asn?Ala?Phe?Pro?Ala?Tyr?Pro?Leu?Ala?Glu?Gly?Asn?Leu?Ala?Val?Val?Gly

Asp?Gly?Ala?Lys?Ile?Gln?Pro?Pro?Leu?Asp?Glu?Asp?Thr?Lys?Val?Leu?His?Ile?Ile?Glu

Glu?Ile?Ala?Thr?Arg?Ser?Asp?Val?Val?Tyr?Gly?Phe?Ala?Glu?Val?Lys?Tyr?Lys?Gln?Leu

Ala?Pro?Leu?Thr?Pro?Ser?Met?Gln?Ala?Asp?Thr?Ala?Ile?Lys?Pro?Val?Val?Ser?Pro?Glu

Val?Val?Asp?Thr?Pro?Glu?Ala?Asn?Asp?Thr?Gly?Leu?Thr?Val?Asp?Ala?Ile?Phe?Thr?Leu

Ser?Glu?Glu?Ala?Leu?Val?Leu?Tyr?Val?Lys?Ala?Leu?Ser?Leu?Leu?Ala?Lys?Ser?Met?Asp

Ile?Ala?Gly?Ala?Trp?Trp?Ala?Arg?Lys?Asn?Arg?Gly?Glu?Thr?Ile?Gly?Asn?Ser?Pro?Asn

Val?Asn?Val?Gly?Cys?Arg?Val?Asn?Asn?Val?Val?Gln?Trp?Val?Arg?Asn?Arg?Phe?Asn?Glu

Val?Leu?Gln?Lys?Ala?Glu?Tyr?Ser?Arg?Leu?Lys?Leu?Leu?Asp?Ala?Gln?Arg?Gln?Leu?Pro

Tyr?Asp?His?Ala?His?His?Ala?Ser?Gln?Val?Gly?Arg?Val?Ser?Val?Gly?Ala?Leu?Ala?His

Ser?Ser?Asp?Gln?Val?Val?Ile?Ser?Ser?Gly?Ile?Thr?Ala?Glu?Lys?Leu?Met?Tyr?Asp?Arg

Ala?Leu?Glu?Met?Ser?Arg?Thr?Ala?Ala?Ile?Asn?Glu?Leu?Thr?Gly?Glu?Asp?Leu?Pro?Gly

Cys?Glu?Ile?Ala?Tyr?Met?Thr?Ala?Ile?Arg?Met?Leu?Glu?Ala?Val?Leu?Asp?Ser?Asp?Glu

Asp?His?Gln?Leu?Ser?Gly?Glu?Arg?Ala?Gly?Glu?Ala?Ala?Asn?Thr?Val?Ser?Asn?Glu?Glu

Asn?Gly?Glu?Val?Asn?Gly?Leu?Gln?Gly?Glu?Asp?Arg?Glu?Ile?Val?Ala?Lys?Leu?Val?Ala

Ser?Ile?Arg?Ile?Arg?Leu?Thr?Ala?Leu?Lys?Lys?Lys?Ile?Ala?Leu?Met?Thr?Lys?Arg?Thr

Ser?Ala?Pro?Ala?Met?Val?Ser?Pro?Ala?Arg?Thr?Pro?Ala?Cys?Gln?Pro?Pro?Ala?Ala?Ser

Pro?Val?Val?Pro?Gln?Ala?Ser?Ser?Arg

SEQ?ID?NO：8

Met?Ala?Thr?Pro?Ser?Asn?Pro?Tyr?Ala?Pro?Arg?Arg?Ser?Gly?Ala?Ser?Pro?Ser?Ser?Ser

Ala?Ala?Ala?Glu?Gln?Ile?Ile?Gly?Lys?Phe?Lys?Arg?Met?Asp?His?Ile?Gly?Lys?Gly?Ser

Phe?Ala?Glu?Val?Tyr?Arg?Gly?Ile?His?Ile?Glu?Lys?Arg?Gln?Ser?Val?Ala?Ile?Lys?Ser

Val?Asn?Met?Asn?Lys?Leu?Asn?Lys?Lys?Leu?Lys?Asp?Asn?Leu?Val?Ser?Glu?Ile?Ser?Ile

Leu?Arg?Ser?Leu?His?His?Pro?His?Ile?Val?Ser?Leu?Ile?Asp?Cys?His?Glu?Thr?Pro?Ser

Arg?Met?His?Ile?Ile?Met?Glu?Phe?Cys?Glu?Leu?Gly?Asp?Leu?Ser?Ala?Phe?Ile?Lys?Lys

Arg?Ala?Asp?Leu?Val?Asn?His?Pro?Gln?Thr?Gln?Arg?Met?Ile?Glu?Lys?Tyr?Pro?Asn?Pro

Ala?Val?Gly?Gly?Leu?Asn?Glu?Val?Ile?Val?Arg?His?Phe?Ala?Lys?Gln?Met?Ala?Ser?Ala

Leu?Glu?Phe?Leu?Arg?Ser?Lys?Asn?Tyr?Ile?His?Arg?Asp?Leu?Lys?Pro?Gln?Asn?Leu?Leu

Leu?Asn?Pro?Ser?Ser?Val?Tyr?Tyr?Ser?Gln?Ser?Gly?Thr?Leu?Glu?Arg?Met?Pro?Leu?Ala

Ala?Asp?Ala?Ser?Ser?Leu?Leu?Pro?Ala?Thr?Gly?Ile?Glu?Ser?Leu?Pro?Met?Leu?Lys?Ile

Ala?Asp?Phe?Gly?Phe?Ala?Arg?Ile?Leu?Pro?Thr?Thr?Ser?Leu?Ala?Glu?Thr?Leu?Cys?Gly

Ser?Pro?Leu?Tyr?Met?Ala?Pro?Glu?Ile?Leu?Arg?Tyr?Glu?Lys?Tyr?Asp?Ala?Lys?Ala?Asp

Leu?Trp?Ser?Val?Gly?Thr?Val?Leu?Phe?Glu?Met?Met?Cys?Ala?Arg?Pro?Pro?Phe?Arg?Ala

Asn?Asn?His?Val?Glu?Leu?Leu?Arg?Lys?Ile?Glu?Glu?Arg?Lys?Asp?His?Ile?Arg?Phe?Pro

Glu?Gly?Ile?Val?Cys?Ser?Arg?Ala?Met?Lys?Asn?Leu?Ile?Arg?Ala?Leu?Leu?Lys?Arg?Lys

Pro?Thr?Glu?Arg?Met?Ser?Tyr?Asp?Ser?Phe?Phe?Ser?Asp?Pro?Val?Ile?Arg?Glu?Glu?Ile

Pro?Asp?Met?Val?Asp?Glu?Asp?Leu?Pro?Gln?Ala?Met?Gln?Ala?Ser?Glu?Pro?Glu?Pro?Pro

Val?Glu?Pro?Pro?Lys?Arg?Val?Gln?Lys?Met?Pro?Val?Glu?Met?Asp?Arg?Arg?Pro?Ser?Asp

Ser?Pro?Tyr?Ser?Arg?Ser?Pro?Arg?Asp?Arg?Thr?Gly?Met?Gly?Ser?Thr?Pro?Pro?Ser?Arg

Pro?Met?Ser?Arg?Pro?Ser?Ser?Ala?Gln?Ala?Ala?Gly?Thr?Pro?Pro?Arg?Thr?Leu?Ser?Met

Arg?Arg?Gly?Ser?Asn?Ala?Pro?Ile?Glu?Pro?Ile?Glu?Glu?His?Ala?Pro?Leu?Arg?Glu?Gln

Arg?Arg?Pro?Val?Leu?Thr?Asn?Ala?Ala?Thr?Ala?Pro?Ala?Arg?Gln?Met?Pro?Leu?Ser?Glu

Gln?Ala?Met?Ala?Gly?His?Arg?Arg?Arg?Tyr?Ser?Arg?Asp?Asp?Gln?Ala?Pro?Val?Pro?Ser

Ser?Leu?Lys?Asp?Thr?Glu?Arg?Glu?Arg?Arg?Thr?Glu?Ala?Gly?Gly?Met?Arg?Glu?Ala?Ala

Glu?Arg?Ala?Ala?Gln?Asp?Ala?Ala?Leu?Asp?Asp?Gly?Phe?Val?Phe?Val?Glu?Lys?Arg?Arg

Val?Glu?Ile?Asp?Ala?Leu?Ala?Asp?Glu?Ile?Ala?Val?Gly?Ser?Pro?Gln?Thr?Gln?Arg?Asp

Arg?Val?Ser?Gln?Arg?Asp?Thr?Met?Lys?Arg?Arg?Ser?Thr?Thr?Gln?Gly?Ala?Pro?Thr?Ser

Thr?Thr?Gly?Ala?Thr?Ala?Pro?Ser?Lys?Ala?Ile?Gln?Ile?Gln?Arg?Gln?Pro?Ser?Leu?Thr

His?Gln?Arg?Ala?Gly?Ser?Tyr?Glu?Arg?Arg?Arg?Pro?Tyr?Arg?Pro?Ser?Phe?Glu?Ser?Ala

Thr?Ser?Ala?Leu?Thr?Lys?Ala?Met?Asn?Met?Val?Ser?Ile?Arg?Gly?Leu?Gly?Leu?Ser?Pro

Pro?Val?Met?Lys?Gly?Val?Ser?Pro?Pro?Gln?Gly?Tyr?Ser?Ala?Phe?Pro?Thr?Tyr?Pro?Ala

Ala?Gln?Ser?Ser?Leu?Leu?Leu?Val?Gly?Asp?Asn?Asp?Gln?Val?Thr?Thr?Lys?Asp?Glu?Ala

Ala?Thr?Thr?Val?Lys?Lys?Ile?Glu?Glu?Leu?Ala?Gln?Leu?Ser?Tyr?Val?Ile?Tyr?Gly?Phe

Ala?Glu?Val?Lys?Tyr?Lys?Gln?Leu?Val?Pro?Val?Ala?Pro?Ser?Asp?Glu?Gly?Leu?Gly?Ile

Gly?Pro?Gly?Gly?Val?Arg?Arg?Lys?Pro?Ser?Thr?Asp?Val?Ile?Glu?Asp?Asp?Asp?Asp?Asp

Asp?Leu?Thr?Pro?Asp?Thr?Ile?Val?Thr?Ile?Ala?Glu?Glu?Ala?Leu?Val?Leu?Tyr?Val?Lys

Thr?Leu?Ala?Leu?Leu?Asn?Lys?Ser?Met?Asp?Val?Ala?Gly?Ser?Tyr?Trp?Asn?Lys?His?Arg

Arg?Gly?Ser?Leu?Ser?Pro?Glu?Ala?Ala?Ser?Arg?Ala?Ala?Val?Ile?Ala?Glu?Arg?Leu?Asn

Arg?Val?Val?Ser?Trp?Val?Arg?Asp?Arg?Phe?Asn?Glu?Cys?Cys?Val?Lys?Ser?Glu?Ile?Ile

Ala?Arg?Lys?Leu?Lys?Gln?Ala?Gln?Gln?His?Leu?Pro?Thr?Asp?His?Pro?Ser?His?Pro?Gln

Asn?Leu?Ser?Ala?Ala?Ser?Gly?Ser?Ala?Thr?Thr?Val?Gly?Ser?Ala?Glu?Asn?Ile?Leu?Leu

Thr?Thr?Gly?Ile?Thr?Ala?Glu?Lys?Leu?Met?Tyr?Asp?Arg?Ala?Ile?Glu?Met?Ser?Arg?Ser

Ala?Ala?Val?Asp?Glu?Leu?Thr?Gly?His?Asn?Leu?Pro?Ser?Cys?Asp?Ile?Asn?Tyr?Ser?Thr

Ala?Ala?Ala?Met?Leu?Glu?Ala?Ile?Leu?Glu?Asp?Glu?Glu?Glu?Ser?Gly?Ser?Arg?Lys?Leu

Asp?Thr?Glu?Glu?Ile?Asn?Gly?Leu?Glu?Thr?Glu?Asp?Arg?Gln?Ser?Ile?Gln?Arg?Leu?Leu

Glu?Glu?Ile?Gln?Arg?Arg?His?Lys?Thr?Leu?Lys?Lys?Lys?Ile?Glu?Ile?Gln?Lys?Ala?Gln

Lys?Arg?Asn?Ser?Ile?Thr?Ssr?Ala?Pro?Ala?Gly?Leu?pro?Ser?Ser?Arg?Gly?Ser?Pro?Ser

Ser?Gln?Gly?Thr?Ala?Arg

SEQ?ID?NO：9

Met?Ala?Asp?Arg?Leu?Pro?Thr?Ser?Thr?Ser?Ser?Gly?Arg?Arg?Arg?Arg?Asp?Gly?Asp?Ala

Ser?Ile?Gly?Glu?Phe?Val?Ile?Gly?Gly?Glu?Ile?Gly?Lys?Gly?Ser?Phe?Ala?Gln?Val?Tyr

Ser?Gly?His?His?Lys?Asn?Ser?Lys?Ala?Ala?Val?Ala?Ile?Lys?Ssr?Val?Glu?Met?Gly?Arg

Leu?Asn?Asn?Lys?Leu?Arg?Glu?Asn?Leu?Tyr?Gly?Glu?Ile?Gln?Ile?Leu?Lys?Thr?Leu?Arg

His?Pro?His?Ile?Val?Ala?Leu?His?Asp?Cys?Val?Glu?Ser?Ala?Thr?His?Ile?Asn?Leu?Val

Met?Glu?Tyr?Cys?Glu?Leu?Gly?Asp?Leu?Ser?Phe?Phe?Ile?Lys?Lys?Arg?Asp?Arg?His?Gly

Thr?Asn?Ala?Ala?Thr?Glu?Asp?Met?Ala?Arg?Lys?Tyr?Pro?Val?Thr?Pro?Gly?Ser?Gly?Leu

His?Glu?Val?Val?Thr?Arg?His?Phe?Leu?Gln?Gln?Leu?Ala?Ser?Ala?Leu?Lys?Phe?Leu?Arg

Glu?Lys?Asn?Tyr?Val?His?Arg?Asp?Val?Lys?Pro?Gln?Asn?Leu?Leu?Leu?Leu?Pro?Ser?Pro

Gly?Phe?Arg?Lys?Glu?Asn?Ser?Arg?Pro?Ile?Leu?Thr?Ala?Ser?Asn?Asp?Ser?Leu?Ile?Pro

Asn?Ala?Gly?Leu?Ala?Ser?Leu?Pro?Met?Leu?Lys?Leu?Ala?Asp?Phe?Gly?Phe?Ala?Arg?Val

Leu?Pro?Ser?Thr?Ser?Leu?Ala?Asp?Thr?Leu?Cys?Gly?Ser?Pro?Leu?Tyr?Met?Ala?Pro?Glu

Ile?Leu?Arg?Tyr?Glu?Arg?Tyr?Asp?Ala?Lys?Ala?Asp?Leu?Trp?Ser?Val?Gly?Thr?Val?Leu

Tyr?Glu?Met?Ile?Thr?Gly?Arg?Pro?pro?Phe?Arg?Ala?Arg?Asn?His?Val?Glu?Leu?Leu?Arg

Lys?Ile?Glu?Ala?Thr?Glu?Asp?Lys?Val?Lys?Tyr?Pro?Lys?Asp?Ala?Val?Val?Ser?Lys?Asp

Leu?Val?Lys?Leu?Ile?Gly?Lys?Leu?Leu?Thr?Arg?Asn?Pro?Val?Glu?Arg?Met?Arg?Phe?Glu

Asp?Phe?Phe?Asn?Asp?Pro?Val?Val?Val?Gly?Pro?Ile?Pro?Gly?Val?Val?Glu?Asp?Asp?Ile

Pro?Lys?Val?Glu?Gln?Lys?Pro?Ser?Arg?Asp?Leu?Arg?Ser?Leu?Glu?Ala?Asp?Pro?Gln?Arg

Glu?Gln?Ssr?Glu?Leu?Ala?Lys?Ser?Pro?Arg?Glu?Arg?Pro?Leu?Arg?Ser?Pro?Gln?Leu?Pro

Ser?Pro?Asp?Glu?Val?Arg?Val?Pro?Gln?Ala?Asn?Val?Ser?Ala?Arg?Thr?Gly?Gln?Ser?Pro

Gly?Arg?Glu?Ile?Gly?Glu?Gly?Leu?Gly?Ile?Arg?Arg?Pro?Pro?Met?Pro?Gln?Pro?Ser?Thr

Ser?Ala?Pro?Ser?Arg?Pro?His?Arg?Leu?Ser?Asn?Ala?Ser?Leu?Asn?Arg?Pro?Pro?Ile?Arg

Ala?Ser?Ala?Ser?Pro?Pro?Thr?Ser?Tyr?Leu?Asn?Glu?Arg?Lys?Leu?Arg?Pro?Val?Thr?Glu

Arg?Ser?Met?Thr?Glu?Gln?Asp?Lys?Ala?Ala?Gln?Asp?Val?Ala?Phe?Glu?Arg?Asp?Tyr?Val

Val?Val?Glu?Lys?Lys?His?Val?Glu?Val?Asn?Ala?Phe?Ala?Asp?Glu?Met?Ala?Ala?Asn?Pro

Arg?Leu?Thr?Ser?Leu?Ser?Pro?Lys?Asn?Gly?Gln?Met?Val?Arg?Arg?Ala?Thr?Gln?Gln?Gly

Pro?Pro?Thr?Ser?Thr?Thr?Gly?Ala?Gly?Arg?Met?Gln?Pro?Ser?Ser?Ala?Val?Gln?Ile?Ala

Gln?Gly?Lys?Gly?Arg?Pro?Gly?His?Asp?His?Pro?Cys?Val?Ser?Leu?Ala?Ser?Arg?Ser?Leu

Asn?Thr?Ser?Ser?Ala?Ala?Arg?Ala?Pro?Leu?Arg?Pro?Cys?Thr?Thr?Arg?Ser?Pro?Arg?Ile

Arg?His?Arg?His?Pro?Leu?Leu?Leu?Leu?Asp?Ser?Ser?Val?Thr?Ala?Asp?Arg?Ala?His?His

Leu?Thr?Lys?Met?His?Gly?Ser?Ser?Ser?Ser?Ser?Lys?Thr?Ser?His?Ile?Val?Val?Ile?Val

Tyr?Met?Ala?Leu?Pro?Lys?Ser?Ser?Leu?Ser?Asn?Phe?Ser?Leu?Trp?His?Leu?Pro?Trp?Ile

Thr?Pro?Trp?Val?Ala?Pro?Leu?Pro?Thr?Lys?Ser?Met?Arg?Met?Val?Leu?Leu?Ser?Arg?Gln

Arg?Trp?Leu?Cys?Leu?Arg?Arg?Leu?Ser?Cys?Phe?Thr?Ser?Arg?Leu

SEQ?ID?NO：10

Met?Ser?Lys?Ser?Ser?Thr?Gly?Arg?Asp?Glu?Arg?Ile?Gly?Asp?Phe?Val?Ile?Glu?Asn?Glu

Ile?Gly?Lys?Gly?Ser?Phe?Ala?Val?Val?His?Lys?Gly?Tyr?Arg?Leu?Gln?Pro?Arg?Glu?Pro

Val?Ala?Ile?Lys?Ile?Val?Ile?Arg?Lys?Lys?Leu?Thr?Pro?Lys?Leu?Leu?Asp?Asn?Leu?Glu

Gly?Glu?Ile?Ala?Ile?Leu?Lys?Ala?Ile?His?His?Pro?Asn?Ile?Val?Glu?Leu?Lys?Glu?Cys

Leu?Lys?Thr?Glu?His?Gln?Ile?Tyr?Leu?Val?Met?Ala?Phe?Cys?Ala?Ser?Gly?Asp?Leu?Ala

Gln?Tyr?Ile?Lys?Lys?Arg?Phe?Asp?Ile?Tyr?Glu?Arg?Ala?Gly?Met?Ala?Glu?Pro?Asp?Ser

Leu?Thr?Lys?Gly?Phe?Lys?Pro?Thr?Tyr?Pro?His?Pro?Val?Asp?Gly?Gly?Leu?Asn?Glu?Thr

Ile?Val?Arg?Ser?Ile?Leu?Thr?Gln?Leu?Ala?Ala?Ala?Leu?Glu?Phe?Met?Arg?Ala?Arg?Asp

Ile?Val?His?Arg?Asp?Ile?Lys?Pro?Gln?Asn?Leu?Leu?Leu?Gln?Pro?Pro?Asp?Ala?Ala?Phe

Leu?Ala?Leu?Gly?Asn?Pro?Arg?Glu?Ile?Pro?Gln?Met?Lys?Val?Ala?Asp?Phe?Gly?Phe?Ala

Arg?His?Leu?Ser?Val?Asn?Thr?Leu?Ala?Glu?Thr?Leu?Cys?Gly?Ser?Pro?Leu?Tyr?Met?Ala

Pro?Glu?Ile?Leu?Arg?Phe?Glu?Lys?Tyr?Asp?Ala?Lys?Ala?Asp?Leu?Trp?Ser?Val?Gly?Ala

Val?Leu?Phe?Glu?Met?Thr?Val?Gly?Lys?Pro?pro?Phe?Arg?Ala?Ala?Asn?His?Val?Glu?Leu

Leu?Lys?Arg?Ile?Glu?Arg?Gly?Glu?Asp?Lys?Ile?Lys?Phe?Pro?Asp?Glu?Arg?Ser?Ala?Gly

Ser?Leu?Ala?Arg?Glu?Ala?Ala?Arg?Arg?Gln?Glu?Leu?Gly?Glu?Ala?Pro?Leu?Pro?Pro?Pro

His?Pro?Val?Ser?Glu?Asp?Val?Lys?Ile?Leu?Ile?Arg?Gln?Leu?Leu?Arg?Gln?Arg?Pro?Val

Ser?Arg?Met?Ser?Phe?Asp?Asp?Phe?Phe?Ala?Ser?Pro?Val?Ile?Ser?Asp?Phe?Lys?Ala?Phe

Ile?Arg?Pro?Arg?Ala?Gln?Pro?Glu?Ala?Val?Glu?Arg?Tyr?Glu?Asp?Leu?Gln?Arg?Ser?Glu

Arg?Ser?Val?Ile?Ile?Pro?Ser?Ser?Gly?Ile?Lys?His?Val?Ser?Val?Ser?Ser?Ile?Glu?Ala

Ser?Thr?Gln?Gln?Pro?Gly?Val?Gln?Pro?Pro?Val?Ser?Thr?Ala?Thr?Ser?Pro?Pro?Ala?Leu

Glu?Ser?Arg?Ser?Thr?Gln?Glu?Ala?Ser?Pro?Lys?Ala?Ile?Thr?Gly?Glu?Thr?Ile?Ala?Pro

Asn?Lys?Thr?Pro?Arg?Glu?Asp?Ala?Arg?Pro?pro?Arg?Thr?Leu?Pro?Arg?Ala?Phe?Ser?Ala

Lys?Tyr?Val?Thr?Gly?Glu?Pro?Pro?Gln?Pro?Glu?Asp?Leu?Glu?Lys?Arg?Val?Pro?Pro?Thr

Met?Thr?Arg?Thr?Pro?Ser?Ser?Pro?Gly?Ile?Pro?Glu?Gly?Ser?Leu?Leu?Ser?Gly?Glu?Arg

Asp?Glu?Ala?Pro?Gln?Ala?Thr?Thr?Glu?His?Phe?Gly?Ser?Ser?Lys?Gly?Gly?Glu?Asp?Ser

Phe?Leu?Gly?Lys?Glu?Tyr?Val?Leu?Ile?Glu?Lys?Gln?Ser?Val?Glu?Val?Asn?Ala?Leu?Ala

Asp?Glu?Leu?Ala?Ala?Ser?Pro?Gln?Ser?Arg?Leu?Gly?Leu?Ala?Ser?Arg?Arg?Pro?Ser?Arg

Leu?Ser?Arg?Leu?Ser?Ser?Gly?Pro?Leu?Pro?Ser?Ala?Pro?Gly?Ala?Ser?Pro?Pro?Thr?Ala

Pro?Pro?Thr?Ile?Leu?Ser?Ser?Lys?Pro?Ile?Arg?Ile?Ala?Asn?Asn?Thr?Asn?Thr?Ala?Ser

Thr?Gly?Ala?Phe?Ala?Leu?Pro?Pro?Gly?Ser?Arg?Pro?Ser?Ser?Phe?Pro?Arg?Arg?Ala?Ser

Leu?Ser?Ser?Ser?Gly?Ser?Pro?Ser?Thr?Arg?Gln?Gly?Gly?Gln?Val?Ile?Thr?Asn?Met?Asp

Ala?Val?Ala?Ser?Thr?Gln?Ser?Asn?Arg?Arg?Asp?Gly?Asn?Ala?Ser?Ser?Phe?Pro?Lys?Asp

Glu?Val?Ser?Val?Leu?Gly?Gln?Arg?Leu?Ala?Gly?Phe?Gly?Leu?Ser?Gly?Ser?Gly?Val?Gly

Gly?Gly?Pro?Ser?Ser?Ala?Leu?Ala?Lys?Ala?Ile?Ser?Met?Ala?Ser?Leu?Arg?Leu?Phe?Gly

Val?Pro?Ser?Gly?Val?Ser?Leu?Arg?Asp?Ala?Ala?Ala?Leu?Val?Arg?Thr?Arg?Ala?Gln?Arg

Arg?Gly?Ile?Ala?Arg?Ala?Thr?Asp?Ser?Leu?Asp?Glu?Ala?Glu?Met?Thr?Leu?Leu?Ser?Thr

Leu?Glu?Asp?Leu?Gly?Gln?Lys?Ala?Phe?Val?Leu?Ser?Glu?Phe?Ala?Asp?Ser?Lys?Leu?Ala

His?Phe?Phe?Pro?Asp?Gly?Pro?His?Gln?Leu?Ser?Gln?Glu?Leu?Asp?Ser?Ser?Thr?Ala?Thr

Ser?Gly?Ile?Ser?Pro?Ser?Arg?Asn?Ser?Val?Gln?Gly?Ser?Ala?Arg?Arg?Val?Gly?Ser?Ile

Ser?Ser?Ser?Ser?Ser?Ser?Ala?Val?Asp?Pro?Val?Ala?Ala?Glu?Ala?Ala?Ser?Ala?Glu?Ala

Leu?Met?Leu?Tyr?Val?Arg?Ser?Leu?Ala?Phe?Leu?Gln?Arg?Ala?Ile?Thr?Leu?Thr?Lys?Arg

His?Val?Glu?Ser?Arg?Ser?Arg?Pro?Gly?Val?Pro?Ala?Val?Thr?Ser?Ala?Glu?Leu?Asn?Asp

Val?Val?Gln?Trp?Leu?Arg?Ala?Arg?Phe?Asn?Glu?Val?Tyr?Asp?Lys?Ala?Asp?Phe?Ala?Arg

Ser?Arg?Cys?Ser?Glu?Leu?Pro?Glu?Ser?Ala?Gln?Gln?Val?Asp?Lys?Leu?Ile?Phe?Asp?Lys

Ala?Val?Glu?Val?Ala?Arg?Ala?Ala?Ala?Thr?Asp?Glu?Leu?Glu?Asn?Asn?Arg?Glu?Gly?Ser

Gly?Trp?Asp?Pro?Ser?His?Cys?Leu?Leu?Ala?Tyr?Glu?Thr?Ala?Asn?Ser?Met?Leu?Ser?Ser

Leu?Leu?Asp?Pro?Gly?Glu?Asp?Ala?Met?Ser?Leu?Ser?Glu?Gly?Ser?Ile?Leu?Met?Ile?Asp

Gly?Tyr?Val?Lys?Ser?Ile?Asn?Lys?Arg?Leu?Trp?Thr?Leu?Gln?Glu?Gln?Phe?Gly?Gly?Gly

Val?Gly?Ala?Val?Gly?Ala?Ala?Gly?Ala?Ser?Pro?Val?Gly?Val?Asp?Ala?Glu?Ala?Arg?Pro

Gly?Val?Ser?Arg?Ser?Arg?Thr?Glu?Ser?Pro

Claims (7)

1, a kind of epiphyte pathogenic gene mgATG1 that comes from Pyricularia oryzae is characterized in that: the nucleotide sequence of this gene or its complementary nucleotide sequence are SEQ ID NO:1.

2, the cDNA sequence of gene mgATG1 coding as claimed in claim 1, it is characterized in that: described cDNA sequence has the nucleotide sequence shown in the SEQID NO:2.

3, gene mgATG1 encoded protein matter as claimed in claim 1, it is characterized in that: described protein has the aminoacid sequence shown in the SEQ IDNO:3.

4, the promotor of gene mgATG1 as claimed in claim 1 is characterized in that: described promotor has the nucleotide sequence shown in the SEQ ID NO:4.

5, the expression that utilizes gene mgATG1 as claimed in claim 1 is used in design and screening antifungal drug as target.

6, utilize protein expression as claimed in claim 3 and modification as target, in design and screening antifungal drug, use.

7, in conjunction with utilizing protein as claimed in claim 3 and promotor as claimed in claim 4, in design and screening antifungal drug, use as target.

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