CN109402065A

CN109402065A - 一种番鸭c亚型禽偏肺病毒弱毒株及其制备和应用

Info

Publication number: CN109402065A
Application number: CN201810897411.3A
Authority: CN
Inventors: 刘佳佳; 陈�峰; 王占新; 鲁俊鹏; 覃健萍; 操胜
Original assignee: Wens Foodstuff Group Co Ltd
Current assignee: Wens Foodstuff Group Co Ltd
Priority date: 2018-08-08
Filing date: 2018-08-08
Publication date: 2019-03-01
Anticipated expiration: 2038-08-08
Also published as: CN109402065B

Abstract

本发明属于微生物动物病毒领域，具体涉及一种番鸭C亚型禽偏肺病毒弱毒株及其制备和应用。本发明的番鸭C亚型禽偏肺病毒弱毒株，保藏号为：CCTCC NO:V201823，核苷酸序列为SEQ ID NO：1，氨基酸序列为SEQ ID NO：2至SEQ ID NO：9。本发明的番鸭C亚型禽偏肺病毒弱毒株，通过将C亚型禽偏肺病毒S‑01株进行传代培养至50代制得。本发明首次获得番鸭C亚型禽偏肺病毒S‑01株致弱培养株，为弱毒苗的研制打下基础。

Description

一种番鸭C亚型禽偏肺病毒弱毒株及其制备和应用

技术领域

本发明属于微生物动物病毒领域，具体涉及一种番鸭C亚型禽偏肺病毒弱毒株及其制备和应用。

背景技术

禽偏肺病是由禽偏肺病毒(avian metapneumovirus，aMPV)引起的上呼吸道感染的急性高度传染性疾病之一。禽肺病最开始在幼龄的火鸡中分离到，伴随着打喷嚏，气管啰音，鼻和眼中有黏着的分泌物，结膜炎等临床症状。因此称之为火鸡鼻气管炎。在1978年的南非，首次分离到aMPV，随后在欧洲，亚洲和南美也有报道。我国于1998 年在有肿头综合征的鸡群中首次报道分离到禽肺病毒A亚型，2012 年又陆续的在鸡群中分离到B、C亚型。2010年7月份在广东省某鸭场发生的以雏鸭、产蛋种鸭咳嗽，产蛋种鸭产蛋率异常下降为特征的鸭群中，分离到一株C亚型偏肺病毒，命名为S-01株，鸭源C亚型禽偏肺病毒仅仅在法国、加拿大有报道。

目前用于aMPV的预防最有效的方法就是疫苗的使用，对于已经感染了的家禽，除了应用不同的抗生素降低发病程度和继发感染外，没有更有效的治疗方法。提高生物安全性及加强饲养环境的管理对该病的发生很重要，接种疫苗可以大大的降低发病率，可以减少经济损失。禽偏肺病毒以细胞免疫为主，目前，已经研发了很多针对aMPV 的疫苗(弱毒疫苗和灭活苗)，但仅是针对于aMPV-A和aMPV-B 亚型的细胞弱毒活疫苗得到全世界范围内广泛应用。aMPV不同亚型之间存在着交叉保护，尤其是aMPV-A和aMPV-B之间的交叉保护。但是不同aMPV-C株之间，aMPV-A 、aMPV-B株与aMPV-C 株之间交叉保护效果不是很理想。因此需要针对aMPV-C亚型毒株研制出弱毒苗。

鸭源C亚型禽偏肺病毒仅仅在法国、加拿大有报道，针对于番鸭源的aMPV-C亚型的弱毒苗目前还未有报道。

发明内容

鉴于此，有必要针对上述问题提供一种鸭C亚型禽偏肺病毒弱毒株及其制备和应用，本发明首次得到鸭源C亚型禽偏肺病毒弱毒苗，致弱后毒株效价较致弱之前细胞毒价升高明显。

本发明是通过以下技术方案实现的：

一种番鸭C亚型禽偏肺病毒弱毒株，保藏号为：CCTCC NO:V201823；保藏单位：中国典型培养物保藏中心；保藏日期：2018 年08月02日；保藏地址：中国.武汉.武汉大学；核苷酸序列为SEQ ID NO：1所示。

一种番鸭C亚型禽偏肺病毒弱毒株，保藏号为：CCTCC NO:V201823；保藏单位：中国典型培养物保藏中心；保藏日期：2018 年08月02日；保藏地址：中国.武汉.武汉大学；所述弱毒株阅读框包括N基因、P基因、M基因、M2基因、SH基因、F基因、G基因、 L基因，所述阅读框的氨基酸序列分别对应SEQ ID NO：2至SEQ ID NO：9。

一种番鸭C亚型禽偏肺病毒弱毒株的制备方法，包括，将C亚型禽偏肺病毒S-01株进行传代培养代数为40-62代。

进一步的，所述番鸭C亚型禽偏肺病毒弱毒株的制备方法，包括，将C亚型禽偏肺病毒S-01株进行传代培养至50代。

进一步的，所述番鸭C亚型禽偏肺病毒弱毒株的制备方法，具体包括：

用番鸭C亚型禽偏肺病毒S-01株，接种长满单层的非洲绿猴肾细胞(Vero)T25细胞瓶，每瓶接种100倍稀释的病毒液500ul；吸附2小时后，弃去病毒液，加入5ml含2％胎牛血清的DMEM培养液，培养24～96小时，至出现细胞病变；反复冻融3次，吸取病毒液接种下一代；如此操作，连续传代培养至40代，再用有限稀释法连续纯化5次后，继续传代培养。

一种包含上述弱毒株或上述方法所制备的弱毒株的鸭免疫用制品，其中每份免疫制品中弱毒株的含量(即每只每次免疫用量)为≧ 10^2.5TCID₅₀；优选≧10^4.5TCID₅₀。

本发明有益效果：

目前暂无鸭源C亚型禽偏肺病毒弱毒苗的报导，本发明属于首次分离传代致弱培养。

本发明致弱后毒株较稳定，S-01-42P、S-01-52P、S-01-62P都完全致弱，未出现不稳定现象。

为了研制出能有效的防控鸭C亚型禽偏肺病毒的疫苗，本研究将番鸭C亚型禽偏肺病毒S-01株连续的在Vero细胞上传代培养，达到致弱的目的，为弱毒苗的研制打下基础。

C亚型禽偏肺病毒S-01株经过在Vero细胞上的连续的传代，通过不同代次致病性试验，40代后毒株已经致弱。致弱后毒株效价较致弱之前细胞毒价升高明显，由原先的10^3.0TCID₅₀/0.1ml变为 10⁵TCID₅₀/0.1ml以上。本发明致弱后代次(S-01-50P)免疫原性最好，10^2.5TCID₅₀/0.1ml临床保护可以达到100％，不能完全阻止排毒， 10^4.5TCID₅₀/0.1ml临床保护达到100％，而且完全可以阻止排毒。因此通过本研究结果选取S-01-50P作为弱毒疫苗候选株(致弱稳定，前10代，后10代均已经致弱)，10^4.5TCID₅₀/0.1ml既可以达到100％的临床保护率及0％的排毒率。致弱株S-01-50P全基因序列的测序分析表明其与初始分离株S-01-5P相比，存在16个氨基酸的变异，这些氨基酸的变异与毒株毒力的变化有很大的关系。

附图说明

图1为发病鸭只鼻腔。

图2为未发病鸭只鼻腔。

具体实施方式

为了更好地说明本发明所解决的问题、所采用的技术方案和所达到的效果，现结合具体实施例和相关资料进一步阐述。需要说明的是，本发明内容包含但不限于以下实施例及其组合实施方式。

本发明实施例中未注明具体技术或条件的，按照本领域内的文献所描述的技术或条件或者按照产品说明书进行。所用试剂或仪器未注明生产厂商者，均为可以通过市购等途径获得的常规产品。

实施例1鸭C亚型偏肺病毒的传代培养

用实验室分离到的在自然界中引起番鸭发病的鸭C亚型禽偏肺病毒S-01株，接种长满单层的非洲绿猴肾细胞(Vero)T25细胞瓶，每瓶接种100倍稀释500ul病毒液，吸附2小时后，弃去病毒液，加入5ml含2％胎牛血清的DMEM培养液，24～96小时不等，培养至出现细胞病变，反复冻融3次，吸取病毒液接种下一代。连续传代培养至40代，后再连续用有限稀释法纯化5次后，传至62代。

传代培养结果

F3代鸭胚卵黄液接种Vero细胞，F1～F3代无明显病变，F4代在第4天出现细胞病变，悬浮细胞增多，合胞体出现等。F4代效价较低为10³TCID₅₀/0.1ml。随着传代次数的增加，病毒滴度变大，到 F60达到10^5.5TCID₅₀/0.1mL。传代过程中发现随着传代次数的增加细胞出现细胞病变的时间缩短，细胞病变也由原来的悬浮死细胞较少，合胞体形成较多，变为悬浮死细胞变多，合胞体变少。各传代代次效价见表1。

表1 S-01株不同传代代次效价的测定结果

实施例2 C亚型禽偏肺病毒S-01株传代代次致病性试验

挑选将上述S-01株传代代次S-01-5P、S-01-17P、S-01-28P、 S-01-32P、S-01-40P、S-01-50P、S-01-60P、Vero细胞液对照组共9 组，点眼滴鼻感染两周龄雏鸭，感染剂量为10⁴TCID₅₀/ml，免疫后每天观察临床症状并记录每组发病鸭只的数目及严重程度，在免疫后第 5天采集每组的泄殖腔拭子及喉拭子，每5个鸭只的拭子放在一个 15ml的EP管里，进行RT-PCR检测，每组共6个待检测样品。通过观察统临床症状、剖检后病变，分析不同代次对鸭的致病力。(RT-PCR 检测方法参考Ali A,Reynolds DL:A reverse transcription-polymerase chain reaction assay for the detection of avian pneumovirus(Colorado strain).Avian Dis1999,43:600–603.)

实验结果：从临床看F5、F17及F32均出现发病，其余代次未出现发病情况，临床症状主要表现为咳嗽、气管啰音两个症状，剖检典型症状的鸭只可见其鼻腔中有粘液堆积(如图1箭头所指)，阴性对照未见这些症状的出现(图2)。F28未出现发病情况，而F32出现发病情况，说明处于该阶段的病毒致弱情况不稳定。F42及以后的代次均未出现发病情况，说明已经致弱。F5、F17、F32发病率依次降低，分别为73％、33％、13％。各代次免疫后出现排毒情况。各代次排毒及发病率见下表2。

表2 S-01株各代次排毒及发病情况表

实施例3 C亚型禽偏肺病毒S-01株致弱代次疫苗效力实验

把上述结果中已经致弱的代次进行免疫原性及不同效价免疫攻毒试验，试验挑选了F40、F50、F60，具体实施操作为200只1日龄的雏番鸭(未免过aMPV及检测aMPV为阴性)随机分成10组，每组20只。40代、50代、60代每个代次分三个梯度(10^2.5、10^3.5、 10^4.5TCID₅₀/0.1ml)。Vero细胞液0为第10组空白组。雏鸭2w时点眼滴鼻方式进行免疫。免疫后每天观察临床症状，第5天进行泄殖腔拭子与喉拭子的采集，每组随机采集10只鸭只，分别放于2ml的EP 管中。每个EP管中加入300ul的含有2000U的双抗的生理盐水。采集的拭子进行RT-PCR检测。免后3w时实验组全部点眼滴鼻方式进行F5代攻毒，攻毒剂量为10^4.5TCID₅₀。攻毒后每天观察临床症状，攻毒5天进行泄殖腔拭子与喉拭子的采集，每组随机采集10只鸭只，分别放于2ml的EP管中。每个EP管中加入300ul的含有2000U的双抗的生理盐水，采集的拭子进行RT-PCR检测。

实验结果：免疫组各组别免疫后均没有发病的情况，免疫3w后 F5代攻毒，5天采集的拭子检测结果显示只有免疫了10^4.5TCID₅₀组别的鸭只没有排毒情况，其他各个代次的10^3.5及10^2.5均有排毒情况的发生。说明104.5免疫效果最好。空白对照组攻毒后出现咳嗽等临床症状，5天检测到排毒情况，说明对照组成立。排毒统计见表3。

表3效力实验排毒统计表

实施例4、C亚型禽偏肺病毒致弱株S-01-50P基因序列的测定分析

上述C亚型禽偏肺病毒致弱株S-01-50P，进行全基因序列的测定分析，与初始分离株S-01全基因进行比对，分析其核苷酸、氨基酸位点的变异情况。

实验结果：致弱株S-01-50P全基因序列的长度14112bp(不包括 polyA尾)，N蛋白序列(395aa)，P蛋白序列(295aa)，M蛋白序列 (255aa)，F蛋白序列(538aa)，M2蛋白序列(185aa)，SH蛋白序列 (176aa)，G蛋白序列(586aa)，L蛋白序列(2006aa)。通过致弱株 S-01-50P与初始分离病变代次S-01全基因序列的比对分析，发现其同源性相对较高，共有16个氨基酸出现变异。N基因未出现变异；P 基因存在2个氨基酸位点的变异，分别为129Aa R-K、233Aa M-I； M基因存在1个氨基酸的变异136Aa V-D；F基因存在6个氨基酸的变异，分别为60Aa C-R、231Aa V-I、334Aa F-L、453Aa E-K、485Aa E-K、528Aa N-S；基因存在3个氨基酸的变异，分别为60Aa P-L、 65Aa A-V、85Aa T-I；L基因存在4个氨基酸的变异，分别为14Aa T-P、 30Aa I-L、186Aa F-L、1756Aa I-T。这些氨基酸的变化可能与毒株毒力减弱以及病毒复制力变强有关。

以上所述实施例仅表达了本发明的几种实施方式，其描述较为具体和详细，但并不能因此而理解为对本发明专利范围的限制。应当指出的是，对于本领域的普通技术人员来说，在不脱离本发明构思的前提下，还可以做出若干变形和改进，这些都属于本发明的保护范围。因此，本发明专利的保护范围应以所附权利要求为准。

序列表

<110> 广东温氏食品集团股份有限公司

<120> 一种鸭C亚型禽偏肺病毒弱毒株及其制备和应用

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<170> SIPOSequenceListing 1.0

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cgggacaagt gaaaatgtct cttcagggga ttcagcttag tgacttgtcc tataagcatg 60

caatccttaa agaatcacag tacacaataa aaagagatgt agggacaacc actgctgtta 120

ctccatcttc tctgcagagg gaagtatcac tcctatgcgg agagatactg tatgccaagc 180

acacagatta ctcacatgca gctgaagtag gaatgcagta cgtgagcacc acattgggag 240

cagagcgtac acagcagata ctaaagaact caggtagtga agtgcaggca gtattgacca 300

agacatactc ccttggtaaa ggcaaaaaca gcaaagggga ggatttgcaa atgttagaca 360

tacatggggt ggaaaaaagt tgggttgaag aagttgacaa agaagcaagg aaaaccatgg 420

cctcagccac aaaggacaac tcagggccaa taccacaaaa tcaaagacca tcatccccag 480

atgctcctat catattactc tgcataggag cattaatctt cacgaagctg gcatcaacaa 540

tcgaagttgg actggagaca gctgttagaa gggcaaaccg tgtgctgaat gatgcattaa 600

agaggttccc gaggattgac atccccaaaa ttgcgaggtc cttttatgat ctgtttgaac 660

agaaagttta ttacaggagc ttgtttatag agtatggcaa agcccttggg tcttcttcca 720

cgggaagcaa agcagagagc ctgtttgtga acattttcat gcaagcttat ggtgcaggcc 780

aaacaatgct aaggtgggga gtgatcgcaa gatcttccaa caatataatg ttgggccatg 840

tctctgtaca agcagaactc aaacaggtca cggaggtata tgatctagtt agagaaatgg 900

gccctgagtc aggtcttctc cacctgaggc aaagccccaa ggctgggttg ttatcacttg 960

ccaattgtcc aaattttgca agtgttgtgc tagggaatgc ctcaggattg gggatacttg 1020

gtatgtatag aggaagggta ccaaacacag agctgtttgc tgcagcagaa agctatgcaa 1080

gaagcctaaa agaaagcaat aagataaatt tctcatctct tggtctgaca gaagaggaaa 1140

aagaagctgc tgagaacttc ctcaacataa atgaagaagg ccagaatgat tatgagtaat 1200

taaaaatggg acaagtcaaa atgtcctttc ctgaggggaa agatatcttg cttatgggca 1260

atgaagctgc aaaagcagca gaagcttttc aaagatcact aaaaaaagta ggacatagga 1320

ggacacagtc aattgttggg gacaaaataa tcacagtgtc tgaaattgta gaaaagccaa 1380

ccataagtaa gtcaaccaag gtcaccacac cccccgaaag gaaaaatgca tggggcgaaa 1440

aaccagacac cacaaggagc ccaacagaag aagccaaaaa cgaggccacg cttgaggatg 1500

caagccggtt gtacgaggag gtctttgctc ctacgagtga tggcaagact cttgccgaga 1560

aaggaaagga aacaactgaa aaaccaaaaa agaaagtgaa attcaagaat gatgaatcag 1620

gaaaatacac aaaactcgag atggaggcac tagaactgtt atcagacaat gaggatgatg 1680

acgcggaatc atccgtgttg acttttgaag aaaaggacac ctctgctctt agccttgaag 1740

ctaggctgga atctattgac gagaaattaa gcatgatact agggctgctg agaacactca 1800

atgttgccac agctgggccc actgcagcta gagacggcat ccgggatgca atggtagggt 1860

tgagagaaga attaattgct gacatcatca aagaagcaaa agggaaggca gccgagatga 1920

tgaaagagga agcaaagcaa aagtcaaaaa tagggaatgg gagcgtaggc ctaactgaga 1980

aggctaaaga actgaacaag atagtagagg atgagagcac aagcggtgaa tcagaggaag 2040

aagaagagga agaagacgaa gaggaaagca acccagatga tgacctatac tctcttacta 2100

tgtagttaat aaaaaacccg ggacaagtgg aaatggagtc ctatctagta gacacctacc 2160

aaggtgttcc ttacactgct gctgtacaaa ctgatttggt ggagaaagac caactacctg 2220

caaggttaac agtatggttc cctctattcc aaaccaacac acctcctaca gtgctgctag 2280

agcagcttaa gaccctgaca attacaactt tgtacacagc ctcccagaac ggcccaatac 2340

tgaaggtcaa tgcatcagca cagggggctg caatgtcagc attgccaaaa agctttgatg 2400

ttagtgcatc agtagcacta gatgattaca gcaaactaga gtttgacaaa ctgacagtgt 2460

gtgagttaaa agcagtctat ttgacaacaa tgaaacctta tggtatggtc tcaaagtttg 2520

tcaattcagc caaggcagat ggaaagaaaa cacatgattt gattgctctc tgcgacttcc 2580

ttgacctaga gaagggagtt ccagtgacta taccagctta cataaagtct gtgtcaataa 2640

aggagagtga atcagcaact gttgaagctg caattagtgg ggaggcagat caagctataa 2700

ctcaagctag gattgcccca tacgctggct tgatcatgat catgacaatg aacaacccta 2760

aggggatctt caaaaaactg ggtgcaggag ttcaggtaat agtagagtta ggagcatacg 2820

tccaagcaga aagcataagc agaatctgca ggaactggag tcaccagggt acaagatatg 2880

tcctgaagtc aagataaaca tagagaatac acccatctaa tcacagaagt aattccattt 2940

ctagcccact catagttaat tcctggtttg atattattta gaaaaaattg ggacaagtga 3000

aaatgtcttg gaaagtggta ctgctactgg tgttgctagc caccccaaca ggggggctag 3060

aagaaagtta cctggaagaa tcatgcagta ctgttactag aggatatctg agtgttttga 3120

ggacagggtg gtatacaaat gtgttcacac ttgaggttgg agatgtggaa aatctcacat 3180

gtaccgacgg acccagctta ataagaacag agcttgaact gacaaaaaat gcacttgagg 3240

aactcaaaac agtatcagca gatcaattgg caaaggaagc taggataatg tcaccaagaa 3300

aagcccggtt tgttctgggt gccatagcat taggtgtggc aactgccgct gcagtgacag 3360

ctggtgtagc aatagccaag acaattaggc tggaaggaga agtggctgca atcaagggtg 3420

ctctcaggaa aacaaatgag gctgtatcta cattaggaaa tggtgtgaga gtactggcaa 3480

cagctgtgaa tgatctcaag gactttataa gtaaaaaatt gacacctgca ataaacaaga 3540

acaagtgtga catttcagac cttaaaatgg cagtaagctt cggacaatac aatcgaagat 3600

tcctcaatgt ggtaagacag ttttctgaca atgcaggtat tacacctgca atatctctag 3660

atttaatgac tgatgccgag cttgtaagag ctataagtaa catgcccaca tcttcaggac 3720

agatcaatct gatgcttgag aatcgggcaa tggtcagaaa ggaaagggtt tgggatcttg 3780

attggagttt atggtagttc cgtggtctat atggtgcagc ttcctatttt cggtgtgata 3840

gacacaccat gttggaaggt gaaggctgct ccattatgtt cagggaaaga cgggagttat 3900

gcatgtcttt tgcgagagga ccaaggctgg tattgtcaaa atgctggatc cacagtttat 3960

tatccgaatg aggaagactg tgaagtgaga agtgatcatg tcctttgtga cacagcagct 4020

gggataaatg tagcaaagga gtcagaagag tgcaacagaa acatctcaac aacaaagtac 4080

ccctgcaagg taagcacagg gcgtcaccca ataagcatgg tggccttatc accactgggt 4140

gccttggtag catgttatga tggggtgagt tgttccattg gaagcaacaa ggttggaata 4200

atcagacctt tggggaaagg gtgttcatac attagcaatc aagacgctga cactgttaca 4260

attgacaaca cagtgtacca attgagcaaa gtagagggag aacaacacac aatcaaaggg 4320

aagcctgtat ctagcaattt tgacccgata gagtttccta aagaccaatt caacatagcc 4380

ctggatcagg tgtttgaaag tgttgagaag agccagagtc tgatagacca gtcaaacaag 4440

atactggata gcaccaaaaa ggggaacgca ggatttgtca tagtgatagt cctcattgtc 4500

ctgcttatgc tagctgcagt tggtgtgggt atcttctttg tggttaaaaa gaggaaaact 4560

gctcccaaat tcccaatgga aatgagtggt gtgaacaaca aaggatttat cccttaactt 4620

tagttactaa aaaattggga caagtgaaga tgtctcgcaa ggccccctgc aaatatgaag 4680

tacggggcaa gtgcaataga ggcagtgaat gcaaatttaa ccataattac tggagttggc 4740

cagacaggta cctgttacta aggtctaatt acctgctgaa tcaattactg agaaatacgg 4800

acagatcaga cggactatca ttaatctcag gagcagggcg agacgatagg acacaagatt 4860

ttgttttagg gtcaacaaat gtagtccaga actacattga taacaatgag aacataacaa 4920

aagcatcagc ttgttacagc ctgtataata tcataaaaca actgcaagag actgacgtga 4980

ggcaggccag ggacaacaaa gttgatgaca gcaagcatgt cgctctacat aatttagtgt 5040

tgtcatatat ggaaatgagc aaaacccctg catccttgat aaacaacttg aagaagcttc 5100

cgaaggaaaa actaaaaaaa ttggcaaagc tgatcattga gttgtcagca ggagtggaaa 5160

atgactccac agctaccatg caagatagtg caaactctga ttaaatgtgg agagcatggc 5220

ctgatcttcc tgaagatgag attggatgat atggtatgga ctaaaaatga attggtagac 5280

ataatttcca ctgagatagt taaagtgcat gctaatatat tcaaatgtag actagaggat 5340

atagaaatca tttatgttaa aacatttcta agttaataaa aaattgggac aagtcaacat 5400

ggagcccctg aaagtctctg gaagtggagg gatgccgatg aagacaaggc ttaatattat 5460

acttgagaag tcaatcaata aaatcttaat catcttagga ttactattaa ctgcctcaac 5520

tgtgattaca atcatactca cagtggaata tataagagtt gaaaatgaac tgcaactttg 5580

caagatggga gcagaggtgg ccaagacaac ccaagaaaca caaacactac cagtgaagac 5640

aactcctatg ctaaccagca caagatcaac taccatatcc gtcaaaacta gatcagtttc 5700

aaggactacc catcccaccg gtcccagctg ctggagagag gaggaaaagt gccagaatat 5760

cacagctaaa tggtccaatt gttttggcac atctctacct gtaagggtga attgcacagc 5820

actaagagaa ttgtgtgatg agcagctagg caatcacaca acagttcaag catcaaggaa 5880

gtgtacatgc atatatgcat taaattggga ttgtggttat gcttgagaga gaaaccacgt 5940

cagcagactt caatgagatc tacagaaaag gaactcaaat gcaaaaatca attttttagt 6000

tatttaaaaa ccatgagtat gtctgaacag tgtcaaggcc aagaaaaaca actcgagaac 6060

aggcaatcca atgattacaa tcaatcagac aaaggaaaac gggacaagtc aacatggagg 6120

tcaagataga gaatgttggc aagtcacagg agcttaaagt caaagtcaag aattttataa 6180

aaaggtctga ttgcaagaaa aaactttttg ccttgatttt agggctgatc agctttgaca 6240

tcactatgaa tataatgctg tctgtcatgt atgtggagtc aaatgaggcc ctgagctcat 6300

gcagggtcca aggaactccc gctccaagag acaacaggac aaacacagaa aacacagcaa 6360

aggaaacaac actccacaca atgaccacga caaggaacac agaagcgggg gggacaaaaa 6420

ccaccaaacc ccaggctgac gaaagagcaa caagcccaag caagaacccc accatcgggg 6480

cagacaaaca caaaacaaca agagcaacaa cagaggcaga gcaggagaaa caaagcaagc 6540

aaaccacaga gccaggcacc agcaccccga agcacatccc cgcaaggcca agcagcaaat 6600

ccccagccac aacaaaaaca acaacacagc ctacaacacc aacagtcgca aaaggaggca 6660

cagcacccaa gaacagacag acaacaacca aaaagaccga agcggacacc ccgacaacaa 6720

gcagagcaaa gcaaaccaac aaacccacag ggacagaaac aacacccccc agagcaacaa 6780

cagaaacaga caaggacaaa gaagggccaa cacagcacac aaccaaagag cagcccgaga 6840

caacggcagg agggacaaca actccacagc caagaagaac aaccagcagg ccagccccaa 6900

caacaaacac caaagagggg gcagaaacca ccgggaccag aacaaccaag agcacccaaa 6960

caagcgcaag cccaccaaga ccaacaagaa gcacacccag caaaacagca acaggaacca 7020

acaagagagc cacaacaaca aagggaccga acacagcaag cacagacaga agacaacaga 7080

ccaggacaac cccaaagcaa gaccaacaga cccaaaccaa ggcaaagaca accacaaaca 7140

aggcccacgc aaaagccgcc accacaccag agcacaacac agacacaaca gacagcatga 7200

aagaaaactc caaggaggac aaaaccacca gggaccccag cagcaaagca acaaccaagc 7260

aagaaaacac cagcaaagga acaaccgcaa caaacctcgg aaacaacacc gaggcaggcg 7320

cgagaacacc cccaacaacc accccgacaa ggcacaccac ggaaccagcc acaagcacag 7380

cggggggaca caccaaagca agaactacaa gatggaaaag cacagccgcc agacagccca 7440

caagaaacaa cacaacagca gataccaaaa cagcccagag caaacaaaca acaccggcgc 7500

aactaggcaa caacacaaca ccagagaaca caacaccacc ggacaacaaa agcaacagcc 7560

aaacaaacgt tgcaccaact gaagaaatag aaattggctc aagcctctgg cgaagaaggt 7620

atgtttatgg accctgcaga gagaatgctc tggagcaccc aatgaatccg tgtcttaagg 7680

acaacactac atggatttat ttggacaatg gaaggaattt gccagcaggt tattatgata 7740

gtaaaacaga taaaataatt tgctatggaa tctatagagg aaactcatat tgttacgggc 7800

gaatagagtg cacatgtaaa aacggcacag gactgctatc ctattgctgt aattcataca 7860

actggagtta gtccaagagc aaattagtta attaaaaaga aggaccaagt taaaaatgga 7920

tccactaaat gaaggggttg tgaatgtgta cttgcctgat tcttatttga aaggtgtaat 7980

atcttttagt gaaaccaatg cattaggatc ttgccttcta gggaaacact accttaaaaa 8040

ggataacaca tcaaaagtag ctatagagag ccctgttgta gaacacatca gattgagaaa 8100

tgctttttag actaggatca aggagaaaaa cctaagggta gtagaacccg tcaacatgca 8160

aagtgaagtc atgagaaatt cttacacctg tgaattgaat ttactcaaac aacttataac 8220

aagaagtaaa gacataagct cactcaaact tgacatgata tgtgactggt tgcaattaaa 8280

atcaacaccc gaaaatccat ctgtattaaa gtttgtggat gtaaggtgta ttcccgactg 8340

ggtaagcact tggtttagca gctggtacaa tttaaacaaa cttatattag agtttaggcg 8400

tgaagaggta gcctgcacgg ggtcaatcat atgtaaaaca atagggagca taatgtttat 8460

tatttcatcg ctcgggtgtg tcatcaaaag caacaagagc aaaagaatta gctttatgac 8520

atacaatcag gtgctgacat ggaaagatgt catgttaagc aggttcaatg caaatctatg 8580

tgtgtggatt agcaacagtc ttaacaaaaa tcaggaagga ttaggtctta gaagcaatct 8640

acaagggacc ctggttaaca aattatatga gattgttgac tctatgctca gtgtttgcag 8700

caatgagggg ttcacccttg ttaaagaatt tgaagggttc ataatgagtg agattctgaa 8760

aataacagag catgctcagt ttagtacaag gttcagaaac actttgttga atgggttagt 8820

agaccaactg gtaaagatga ggggcctaaa caggaaaagg gtttcaggca ctgtcttgga 8880

aggaaaccag tatccaatgt atgagacaac attagctaca ctaggggaag ctttaaaaac 8940

aataagactc ctggtaaaca agaacttaga caatgccgct gagctgtact acattttcag 9000

gatttttggc catccaatgg ttgaagagag agaagctatg gacgcagtca gacttaataa 9060

tgagatcaca aaaattttga aattagagag ccttacagag ttgagggggg ccttcatttt 9120

aagaatcata aaaggctttg ttgacacaaa taagagatgg cctaaaatca aaaatctcaa 9180

agtgttgagc agaaggtggg ttatgtactt taaagcaaag agttatccaa gccaattgga 9240

gctgagtagt caagatttcc ttgaattagc aggtgtgcag tttgagcagg aatttgcaat 9300

accagagaga acaaatcttg agatggtctt gaatgataag gccatctcac cacccaaaaa 9360

tttaatatgg tcagtgttcc ctaaaaatta ccttccaaca aacatcaggg aaaaattcac 9420

tgatgaaatg ttcaactcaa gtgaaaagtt gaagacaaga agggtgttgg agtactactt 9480

aaaagacaac aagtttgatc agaatgacct gaaaaaatat gttgttaggc aagagtactt 9540

gggtgataaa gaacatgtag tgtcactgac agggaaagaa cgtgagttaa gtgtgggaag 9600

gatgttcgcg atgcagccag ggaaacaaag gcaagttcag atcttagcag aaaagctact 9660

tgcagacaat atagtaccat tcttccctga gacattgaca aaatatggtg atctagaact 9720

acaaaggatt atggaaataa aatctgagtt gtcatcagta aagtcacgga gaaatgacag 9780

ttacaacaat tatattgcta gggcatcaat agtgactgat cttagtaaat tcaaccaagc 9840

ttttcggtac gagacatctt ctgtctgtgc agatgtagtt gatgaattgc atggcacaca 9900

aagtctcttc tgctggctac atttgactgt gccactgaca actatgattt gcacatatag 9960

acatgctcct ccagaaacag aaggagtgta tgacattgat aaaatcaagg aacaaagtgg 10020

gctttacagg tttcatatgg gggggattga agggtggtgc caaaagttgt ggactatgga 10080

ggctatttct ttattagatg tagtatcagt caaaaaccgt gtccaactaa catcattgtt 10140

aaatggagac aatcaatcta ttgatgttag caaacctgtt aggttgtcac aaggagttga 10200

tgaagttaaa gctgactata gcctggcagt gaaaatgtta aaagaaataa gaaatgctta 10260

caaagacata gggcacaagt taaaagaagg agagacctat atatccagag atctacaatt 10320

catgagcaag gtcattcaat cagagggtgt gatgcatccg tcaccaataa agaaaattct 10380

gagggtcgga ccatggatta atactatctt agatgacatc aagaccagtg cagagtcaat 10440

aggaagccta tgtcaagagt tagagtttag aggtgaaagc ctcttggtta gccttattct 10500

aaggaatttt tggttatacg agttatggat gcatgaatct aaaagtcatc cacttgcagg 10560

gaagcagcta tacagacagc tgagcaagac attagcaata actcaaaaat tctttggaat 10620

aacaaaagaa actgatgttg ttaatctgtg gatgaatgtt ccaatgcaat ttggtggggg 10680

tgaccctgta gtgttgtata ggtcatttta ccggaggaca ccagatttct tgactgaagc 10740

agtaagtcat atgagtgtac tactcaaggt gtatgggaaa gcaaaagagg gatcaaagaa 10800

agattttttc aaagcattat tatcagtaga caaaaataag agggccactc tgaccacatt 10860

aatgagagac cctcaagcag tgggatcaga gaggcaggca agagttacta gtgaaattaa 10920

tagagcagca gtcacaagtg tcttgagctt atcacctaac cagcttttct gtgatagtgc 10980

aatacactac agcagaaatg aagaagaagt tgggttaata gcccaaaaca ttacaccagt 11040

ctatccacac gggctgaggg tgttgtatga gtcattgcca ttccacaagg cggaaaaagt 11100

tgtaaacatg atatctggaa caaaatctat cacaaactta ctacaaagga catcagcaat 11160

caatggagaa gatattgata gagcagtctc aatgatgttg gagaatttag ggttactctc 11220

taggatactg tctgtgagcc aagatgatat aatcttgcct accaaggcaa atggtgatct 11280

gatatgttgt caagtctcca ggactttgag ggaaaggtcc tgggataaca tggaaattgt 11340

tggagtcacc tcaccgagca tagttacatg tatgaacata gtgtattcca gcagctctca 11400

actaaaagga ataacaatcg aaaagttcag cacagataaa actacaagag gtcaaagagg 11460

accaaaaagt ccttgggttg gatccagcac acaggagaaa aaattggttc ctgtttacaa 11520

tagacaaatc ctctcaaagc aacagaaaga acaactagag gcaatcggaa aactgagatg 11580

ggtttataag gggactcaag ggttaaggag gttgttggat aaaatctgca tagggagctt 11640

agggatcagt tacaagaatg ttaagccact cttgccaaga tttatgagtg ttaacttcct 11700

acacagactt tccgtcagta gtagaccaat ggagtttcca gcatctgttc ctgcttacag 11760

aacgaccaat taccatttcg acacaagtcc tgtaaaccag actctgagtg aaagatttgg 11820

aaatgaggac atcaatttgg tgtttcagaa cgctatcagc tgtgggataa gtgtgatgag 11880

tgtagtagag caacttacag gcagaagccc taaacagtta gtaatgattc cccaattaga 11940

agaaattgac ataatgcctc cgcctgtctt ttcaggaaaa tttgattata aactggtaga 12000

aaagatatca tcagaccaac acattttcag ccctgacaaa ttggacctag tgactctagg 12060

aaaaatgtta atgccttcaa caagtggagc aaagtctgat cagttttgga acaggaaaga 12120

aaacttcttc catggcaata atttggtaga gtccctatct gcagcattgg cttgtcattg 12180

gtgcgggatc ttaactgaac agtgtaatga gaacaatatt ttcagaagag aatggggtga 12240

tggttttgtt acagatcatg ccttcataga tttcaaaata tttatcggcg tatttaagac 12300

caaactattg tgtggatggg gatccagggg gggggacatt aaagatcgag acatgataga 12360

cgaatctatt gacaaattaa taagagttga caactcattc tggaggatgt ttagcaaagt 12420

gatacttgaa ccaaaagtaa ggaaaagagt aatgcttttt gatgtgaaga tcttatcact 12480

tgtaggctat gcaggattta aaaattggtt tattgaccac ctgagatcta gtgacttgtg 12540

tgaagtgcca tgggttgtca atgctgatag tgaaattgtt gaggtgtctg ctgtgaaaat 12600

atatttacag ctgctgaggg tgagttctcc cttgaggatt acagtgttga attattcaga 12660

catggcccat gctattacca ggctaataag aaggaaatca atgcatgaca atgtgccatc 12720

aatatctagg acactgtcac ctgcagagtt agctcctgta gttgaaccaa ccgtgcagat 12780

gaacttgttc cctaagatca catttgaaag attgaagaat tatgaaacta tatcagggtc 12840

tactagaggg aagttgacaa gaaattacat ggtgatgttg ccttggcagc atattaatag 12900

gttcaacttt gtattcagct ctaccggttg caaaataagt gtcaaatcat gtattgggaa 12960

gttgattcaa gacttaaatc cgacagtttt ttattttgtg ggagaaggag ccggcaattg 13020

gatggcaaga acagcttgtg agtacccaaa tacaaagttt gtttacagga gtcttaaaga 13080

tgatttagac caccatttcc ctttggaatt tcagagagtc ctagggaaca tgaatagagt 13140

tatagatggt ggagaaggtc tatccatgga cacaacagat gcaactcaga aaactcattg 13200

ggatctgaca catagaattt gtaaagatgc attactgatt actttgtgtg atgctgaatt 13260

taaagatagg gatgactttt tcaagatggt gacactatgg agaaagcatg ttttgtcatg 13320

taggatatgt acaacctatg gaacagatct ctacctgttc gcaaaatatc atgcaaaaga 13380

ggaaagcata aaacttccct attttgttag atcaattgcc acatatgtca tgcaaggcag 13440

taaattgtca ggatctgagt gctatgttct tctgacactt ggtcacccac aacaacctac 13500

catgtcacgg ggaagtacag agctccaaat taaaaatggc tgtgtgtaat gacttcagca 13560

ttcctaggaa agtagaagtg aaggctgttg aggcaaactg caaatcgctg ctttctggcc 13620

taaggacacc tataaacaga gcagaattgg atcgacaaaa gaagatgttg acactaagaa 13680

gttatcattc atcagtagca acagttggag gcagcagagt catcgaatca aaatggttga 13740

gcaagaaagc taccactata atcgagtggt tagaacatat cttgaattcc ccaaaagggg 13800

agttaaacta tgattttttt gaggcactag agaacaccta tccaaatatg gttaaattat 13860

tagataatct tggcagtgct gaactaaaaa aactgatcaa agtgacaggt tacatgctga 13920

tgagtaagaa ataataggag aaaattacct taagagatta cttataattg aggctaaaaa 13980

ctaacatatt catgctgaag gatattaaaa tgatagttgt ttaaatttag caatctgaat 14040

atgaagcaca caatcagcaa ttagttatta aaaaatagag aaactaaaat tggatgaata 14100

cggttttttt gc 14112

<210> 2

<211> 394

<212> PRT

<213> 人工序列(Artificial Sequence)

<400> 2

Met Ser Leu Gln Gly Ile Gln Leu Ser Asp Leu Ser Tyr Lys His Ala

1 5 10 15

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

20 25 30

Thr Ala Val Thr Pro Ser Ser Leu Gln Arg Glu Val Ser Leu Leu Cys

35 40 45

Gly Glu Ile Leu Tyr Ala Lys His Thr Asp Tyr Ser His Ala Ala Glu

50 55 60

Val Gly Met Gln Tyr Val Ser Thr Thr Leu Gly Ala Glu Arg Thr Gln

65 70 75 80

Gln Ile Leu Lys Asn Ser Gly Ser Glu Val Gln Ala Val Leu Thr Lys

85 90 95

Thr Tyr Ser Leu Gly Lys Gly Lys Asn Ser Lys Gly Glu Asp Leu Gln

100 105 110

Met Leu Asp Ile His Gly Val Glu Lys Ser Trp Val Glu Glu Val Asp

115 120 125

Lys Glu Ala Arg Lys Thr Met Ala Ser Ala Thr Lys Asp Asn Ser Gly

130 135 140

Pro Ile Pro Gln Asn Gln Arg Pro Ser Ser Pro Asp Ala Pro Ile Ile

145 150 155 160

Leu Leu Cys Ile Gly Ala Leu Ile Phe Thr Lys Leu Ala Ser Thr Ile

165 170 175

Glu Val Gly Leu Glu Thr Ala Val Arg Arg Ala Asn Arg Val Leu Asn

180 185 190

Asp Ala Leu Lys Arg Phe Pro Arg Ile Asp Ile Pro Lys Ile Ala Arg

195 200 205

Ser Phe Tyr Asp Leu Phe Glu Gln Lys Val Tyr Tyr Arg Ser Leu Phe

210 215 220

Ile Glu Tyr Gly Lys Ala Leu Gly Ser Ser Ser Thr Gly Ser Lys Ala

225 230 235 240

Glu Ser Leu Phe Val Asn Ile Phe Met Gln Ala Tyr Gly Ala Gly Gln

245 250 255

Thr Met Leu Arg Trp Gly Val Ile Ala Arg Ser Ser Asn Asn Ile Met

260 265 270

Leu Gly His Val Ser Val Gln Ala Glu Leu Lys Gln Val Thr Glu Val

275 280 285

Tyr Asp Leu Val Arg Glu Met Gly Pro Glu Ser Gly Leu Leu His Leu

290 295 300

Arg Gln Ser Pro Lys Ala Gly Leu Leu Ser Leu Ala Asn Cys Pro Asn

305 310 315 320

Phe Ala Ser Val Val Leu Gly Asn Ala Ser Gly Leu Gly Ile Leu Gly

325 330 335

Met Tyr Arg Gly Arg Val Pro Asn Thr Glu Leu Phe Ala Ala Ala Glu

340 345 350

Ser Tyr Ala Arg Ser Leu Lys Glu Ser Asn Lys Ile Asn Phe Ser Ser

355 360 365

Leu Gly Leu Thr Glu Glu Glu Lys Glu Ala Ala Glu Asn Phe Leu Asn

370 375 380

Ile Asn Glu Glu Gly Gln Asn Asp Tyr Glu

385 390

<210> 3

<211> 294

<212> PRT

<213> 人工序列(Artificial Sequence)

<400> 3

Met Ser Phe Pro Glu Gly Lys Asp Ile Leu Leu Met Gly Asn Glu Ala

1 5 10 15

Ala Lys Ala Ala Glu Ala Phe Gln Arg Ser Leu Lys Lys Val Gly His

20 25 30

Arg Arg Thr Gln Ser Ile Val Gly Asp Lys Ile Ile Thr Val Ser Glu

35 40 45

Ile Val Glu Lys Pro Thr Ile Ser Lys Ser Thr Lys Val Thr Thr Pro

50 55 60

Pro Glu Arg Lys Asn Ala Trp Gly Glu Lys Pro Asp Thr Thr Arg Ser

65 70 75 80

Pro Thr Glu Glu Ala Lys Asn Glu Ala Thr Leu Glu Asp Ala Ser Arg

85 90 95

Leu Tyr Glu Glu Val Phe Ala Pro Thr Ser Asp Gly Lys Thr Leu Ala

100 105 110

Glu Lys Gly Lys Glu Thr Thr Glu Lys Pro Lys Lys Lys Val Lys Phe

115 120 125

Lys Asn Asp Glu Ser Gly Lys Tyr Thr Lys Leu Glu Met Glu Ala Leu

130 135 140

Glu Leu Leu Ser Asp Asn Glu Asp Asp Asp Ala Glu Ser Ser Val Leu

145 150 155 160

Thr Phe Glu Glu Lys Asp Thr Ser Ala Leu Ser Leu Glu Ala Arg Leu

165 170 175

Glu Ser Ile Asp Glu Lys Leu Ser Met Ile Leu Gly Leu Leu Arg Thr

180 185 190

Leu Asn Val Ala Thr Ala Gly Pro Thr Ala Ala Arg Asp Gly Ile Arg

195 200 205

Asp Ala Met Val Gly Leu Arg Glu Glu Leu Ile Ala Asp Ile Ile Lys

210 215 220

Glu Ala Lys Gly Lys Ala Ala Glu Ile Met Lys Glu Glu Ala Lys Gln

225 230 235 240

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

245 250 255

Glu Leu Asn Lys Ile Val Glu Asp Glu Ser Thr Ser Gly Glu Ser Glu

260 265 270

Glu Glu Glu Glu Glu Glu Asp Glu Glu Glu Ser Asn Pro Asp Asp Asp

275 280 285

Leu Tyr Ser Leu Thr Met

290

<210> 4

<211> 254

<212> PRT

<213> 人工序列(Artificial Sequence)

<400> 4

Met Glu Ser Tyr Leu Val Asp Thr Tyr Gln Gly Val Pro Tyr Thr Ala

1 5 10 15

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

20 25 30

Thr Val Trp Phe Pro Leu Phe Gln Thr Asn Thr Pro Pro Thr Val Leu

35 40 45

Leu Glu Gln Leu Lys Thr Leu Thr Ile Thr Thr Leu Tyr Thr Ala Ser

50 55 60

Gln Asn Gly Pro Ile Leu Lys Val Asn Ala Ser Ala Gln Gly Ala Ala

65 70 75 80

Met Ser Ala Leu Pro Lys Ser Phe Asp Val Ser Ala Ser Val Ala Leu

85 90 95

Asp Asp Tyr Ser Lys Leu Glu Phe Asp Lys Leu Thr Val Cys Glu Leu

100 105 110

Lys Ala Val Tyr Leu Thr Thr Met Lys Pro Tyr Gly Met Val Ser Lys

115 120 125

Phe Val Asn Ser Ala Lys Ala Asp Gly Lys Lys Thr His Asp Leu Ile

130 135 140

Ala Leu Cys Asp Phe Leu Asp Leu Glu Lys Gly Val Pro Val Thr Ile

145 150 155 160

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

165 170 175

Val Glu Ala Ala Ile Ser Gly Glu Ala Asp Gln Ala Ile Thr Gln Ala

180 185 190

Arg Ile Ala Pro Tyr Ala Gly Leu Ile Met Ile Met Thr Met Asn Asn

195 200 205

Pro Lys Gly Ile Phe Lys Lys Leu Gly Ala Gly Val Gln Val Ile Val

210 215 220

Glu Leu Gly Ala Tyr Val Gln Ala Glu Ser Ile Ser Arg Ile Cys Arg

225 230 235 240

Asn Trp Ser His Gln Gly Thr Arg Tyr Val Leu Lys Ser Arg

245 250

<210> 5

<211> 184

<212> PRT

<213> 人工序列(Artificial Sequence)

<400> 5

Met Ser Arg Lys Ala Pro Cys Lys Tyr Glu Val Arg Gly Lys Cys Asn

1 5 10 15

Arg Gly Ser Glu Cys Lys Phe Asn His Asn Tyr Trp Ser Trp Pro Asp

20 25 30

Arg Tyr Leu Leu Leu Arg Ser Asn Tyr Leu Leu Asn Gln Leu Leu Arg

35 40 45

Asn Thr Asp Arg Ser Asp Gly Leu Ser Leu Ile Ser Gly Ala Gly Arg

50 55 60

Asp Asp Arg Thr Gln Asp Phe Val Leu Gly Ser Thr Asn Val Val Gln

65 70 75 80

Asn Tyr Ile Asp Asn Asn Glu Asn Ile Thr Lys Ala Ser Ala Cys Tyr

85 90 95

Ser Leu Tyr Asn Ile Ile Lys Gln Leu Gln Glu Thr Asp Val Arg Gln

100 105 110

Ala Arg Asp Asn Lys Val Asp Asp Ser Lys His Val Ala Leu His Asn

115 120 125

Leu Val Leu Ser Tyr Met Glu Met Ser Lys Thr Pro Ala Ser Leu Ile

130 135 140

Asn Asn Leu Lys Lys Leu Pro Lys Glu Lys Leu Lys Lys Leu Ala Lys

145 150 155 160

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

165 170 175

Met Gln Asp Ser Ala Asn Ser Asp

180

<210> 6

<211> 175

<212> PRT

<213> 人工序列(Artificial Sequence)

<400> 6

Met Glu Pro Leu Lys Val Ser Gly Ser Gly Gly Met Pro Met Lys Thr

1 5 10 15

Arg Leu Asn Ile Ile Leu Glu Lys Ser Ile Asn Lys Ile Leu Ile Ile

20 25 30

Leu Gly Leu Leu Leu Thr Ala Ser Thr Val Ile Thr Ile Ile Leu Thr

35 40 45

Val Glu Tyr Ile Arg Val Glu Asn Glu Leu Gln Leu Cys Lys Met Gly

50 55 60

Ala Glu Val Ala Lys Thr Thr Gln Glu Thr Gln Thr Leu Pro Val Lys

65 70 75 80

Thr Thr Pro Met Leu Thr Ser Thr Arg Ser Thr Thr Ile Ser Val Lys

85 90 95

Thr Arg Ser Val Ser Arg Thr Thr His Pro Thr Gly Pro Ser Cys Trp

100 105 110

Arg Glu Glu Glu Lys Cys Gln Asn Ile Thr Ala Lys Trp Ser Asn Cys

115 120 125

Phe Gly Thr Ser Leu Pro Val Arg Val Asn Cys Thr Ala Leu Arg Glu

130 135 140

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

145 150 155 160

Lys Cys Thr Cys Ile Tyr Ala Leu Asn Trp Asp Cys Gly Tyr Ala

165 170 175

<210> 7

<211> 537

<212> PRT

<213> 人工序列(Artificial Sequence)

<400> 7

Met Ser Trp Lys Val Val Leu Leu Leu Val Leu Leu Ala Thr Pro Thr

1 5 10 15

Gly Gly Leu Glu Glu Ser Tyr Leu Glu Glu Ser Cys Ser Thr Val Thr

20 25 30

Arg Gly Tyr Leu Ser Val Leu Arg Thr Gly Trp Tyr Thr Asn Val Phe

35 40 45

Thr Leu Glu Val Gly Asp Val Glu Asn Leu Thr Arg Thr Asp Gly Pro

50 55 60

Ser Leu Ile Arg Thr Glu Leu Glu Leu Thr Lys Asn Ala Leu Glu Glu

65 70 75 80

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

85 90 95

Ser Pro Arg Lys Ala Arg Phe Val Leu Gly Ala Ile Ala Leu Gly Val

100 105 110

Ala Thr Ala Ala Ala Val Thr Ala Gly Val Ala Ile Ala Lys Thr Ile

115 120 125

Arg Leu Glu Gly Glu Val Ala Ala Ile Lys Gly Ala Leu Arg Lys Thr

130 135 140

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

145 150 155 160

Ala Val Asn Asp Leu Lys Asp Phe Ile Ser Lys Lys Leu Thr Pro Ala

165 170 175

Ile Asn Lys Asn Lys Cys Asp Ile Ser Asp Leu Lys Met Ala Val Ser

180 185 190

Phe Gly Gln Tyr Asn Arg Arg Phe Leu Asn Val Val Arg Gln Phe Ser

195 200 205

Asp Asn Ala Gly Ile Thr Pro Ala Ile Ser Leu Asp Leu Met Thr Asp

210 215 220

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

225 230 235 240

Ile Asn Leu Met Leu Glu Asn Arg Ala Met Val Arg Arg Lys Gly Phe

245 250 255

Gly Ile Leu Ile Gly Val Tyr Gly Ser Ser Val Val Tyr Met Val Gln

260 265 270

Leu Pro Ile Phe Gly Val Ile Asp Thr Pro Cys Trp Lys Val Lys Ala

275 280 285

Ala Pro Leu Cys Ser Gly Lys Asp Gly Ser Tyr Ala Cys Leu Leu Arg

290 295 300

Glu Asp Gln Gly Trp Tyr Cys Gln Asn Ala Gly Ser Thr Val Tyr Tyr

305 310 315 320

Pro Asn Glu Glu Asp Cys Glu Val Arg Ser Asp His Val Leu Cys Asp

325 330 335

Thr Ala Ala Gly Ile Asn Val Ala Lys Glu Ser Glu Glu Cys Asn Arg

340 345 350

Asn Ile Ser Thr Thr Lys Tyr Pro Cys Lys Val Ser Thr Gly Arg His

355 360 365

Pro Ile Ser Met Val Ala Leu Ser Pro Leu Gly Ala Leu Val Ala Cys

370 375 380

Tyr Asp Gly Val Ser Cys Ser Ile Gly Ser Asn Lys Val Gly Ile Ile

385 390 395 400

Arg Pro Leu Gly Lys Gly Cys Ser Tyr Ile Ser Asn Gln Asp Ala Asp

405 410 415

Thr Val Thr Ile Asp Asn Thr Val Tyr Gln Leu Ser Lys Val Glu Gly

420 425 430

Glu Gln His Thr Ile Lys Gly Lys Pro Val Ser Ser Asn Phe Asp Pro

435 440 445

Ile Glu Phe Pro Lys Asp Gln Phe Asn Ile Ala Leu Asp Gln Val Phe

450 455 460

Glu Ser Val Glu Lys Ser Gln Ser Leu Ile Asp Gln Ser Asn Lys Ile

465 470 475 480

Leu Asp Ser Thr Lys Lys Gly Asn Ala Gly Phe Val Ile Val Ile Val

485 490 495

Leu Ile Val Leu Leu Met Leu Ala Ala Val Gly Val Gly Ile Phe Phe

500 505 510

Val Val Lys Lys Arg Lys Thr Ala Pro Lys Phe Pro Met Glu Met Ser

515 520 525

Gly Val Asn Asn Lys Gly Phe Ile Pro

530 535

<210> 8

<211> 585

<212> PRT

<213> 人工序列(Artificial Sequence)

<400> 8

Met Glu Val Lys Ile Glu Asn Val Gly Lys Ser Gln Glu Leu Arg Val

1 5 10 15

Lys Val Lys Asn Phe Ile Lys Arg Ser Asp Cys Lys Lys Lys Leu Phe

20 25 30

Ala Leu Ile Leu Gly Leu Ile Ser Phe Asp Ile Thr Met Asn Ile Met

35 40 45

Leu Ser Val Met Tyr Val Glu Ser Asn Glu Ala Leu Ser Ser Cys Arg

50 55 60

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

65 70 75 80

Thr Ala Lys Glu Thr Thr Leu His Thr Met Thr Thr Thr Arg Asn Thr

85 90 95

Glu Ala Gly Gly Thr Lys Thr Thr Lys Pro Gln Ala Asp Glu Arg Ala

100 105 110

Thr Ser Pro Ser Lys Asn Pro Thr Ile Gly Ala Asp Lys His Lys Thr

115 120 125

Thr Arg Ala Thr Thr Glu Ala Glu Gln Glu Lys Gln Ser Lys Gln Thr

130 135 140

Thr Glu Pro Gly Thr Ser Thr Pro Lys His Ile Pro Ala Arg Pro Ser

145 150 155 160

Ser Lys Ser Pro Ala Thr Thr Lys Thr Thr Thr Gln Pro Thr Thr Pro

165 170 175

Thr Val Ala Lys Gly Gly Thr Ala Pro Lys Asn Arg Gln Thr Thr Thr

180 185 190

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

195 200 205

Asn Lys Pro Thr Gly Thr Glu Thr Thr Pro Pro Arg Ala Thr Thr Glu

210 215 220

Thr Asp Lys Asp Lys Glu Gly Pro Thr Gln His Thr Thr Lys Glu Gln

225 230 235 240

Pro Glu Thr Thr Ala Gly Gly Thr Thr Thr Pro Gln Pro Arg Arg Thr

245 250 255

Thr Ser Arg Pro Ala Pro Thr Thr Asn Thr Lys Glu Gly Ala Glu Thr

260 265 270

Thr Gly Thr Arg Thr Thr Lys Ser Thr Gln Thr Ser Ala Ser Pro Pro

275 280 285

Arg Pro Thr Arg Ser Thr Pro Ser Lys Thr Ala Thr Gly Thr Asn Lys

290 295 300

Arg Ala Thr Thr Thr Lys Gly Pro Asn Thr Ala Ser Thr Asp Arg Arg

305 310 315 320

Gln Gln Thr Arg Thr Thr Pro Lys Gln Asp Gln Gln Thr Gln Thr Lys

325 330 335

Ala Lys Thr Thr Thr Asn Lys Ala His Ala Lys Ala Ala Thr Thr Pro

340 345 350

Glu His Asn Thr Asp Thr Thr Asp Ser Met Lys Glu Asn Ser Lys Glu

355 360 365

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

370 375 380

Asn Thr Ser Lys Gly Thr Thr Ala Thr Asn Leu Gly Asn Asn Thr Glu

385 390 395 400

Ala Gly Ala Arg Thr Pro Pro Thr Thr Thr Pro Thr Arg His Thr Thr

405 410 415

Glu Pro Ala Thr Ser Thr Ala Gly Gly His Thr Lys Ala Arg Thr Thr

420 425 430

Arg Trp Lys Ser Thr Ala Ala Arg Gln Pro Thr Arg Asn Asn Thr Thr

435 440 445

Ala Asp Thr Lys Thr Ala Gln Ser Lys Gln Thr Thr Pro Ala Gln Leu

450 455 460

Gly Asn Asn Thr Thr Pro Glu Asn Thr Thr Pro Pro Asp Asn Lys Ser

465 470 475 480

Asn Ser Gln Thr Asn Val Ala Pro Thr Glu Glu Ile Glu Ile Gly Ser

485 490 495

Ser Leu Trp Arg Arg Arg Tyr Val Tyr Gly Pro Cys Arg Glu Asn Ala

500 505 510

Leu Glu His Pro Met Asn Pro Cys Leu Lys Asp Asn Thr Thr Trp Ile

515 520 525

Tyr Leu Asp Asn Gly Arg Asn Leu Pro Ala Gly Tyr Tyr Asp Ser Lys

530 535 540

Thr Asp Lys Ile Ile Cys Tyr Gly Ile Tyr Arg Gly Asn Ser Tyr Cys

545 550 555 560

Tyr Gly Arg Ile Glu Cys Thr Cys Lys Asn Gly Thr Gly Leu Leu Ser

565 570 575

Tyr Cys Cys Asn Ser Tyr Asn Trp Ser

580 585

<210> 9

<211> 2005

<212> PRT

<213> 人工序列(Artificial Sequence)

<400> 9

Met Asp Pro Leu Asn Glu Gly Val Val Asn Val Tyr Leu Pro Asp Ser

1 5 10 15

Tyr Leu Lys Gly Val Ile Ser Phe Ser Glu Thr Asn Ala Leu Gly Ser

20 25 30

Cys Leu Leu Gly Lys His Tyr Leu Lys Lys Asp Asn Thr Ser Lys Val

35 40 45

Ala Ile Glu Ser Pro Val Val Glu His Ile Arg Leu Arg Asn Ala Phe

50 55 60

Gln Thr Arg Ile Lys Glu Lys Asn Leu Arg Val Val Glu Pro Val Asn

65 70 75 80

Met Gln Ser Glu Val Met Arg Asn Ser Tyr Thr Cys Glu Leu Asn Leu

85 90 95

Leu Lys Gln Leu Ile Thr Arg Ser Lys Asp Ile Ser Ser Leu Lys Leu

100 105 110

Asp Met Ile Cys Asp Trp Leu Gln Leu Lys Ser Thr Pro Glu Asn Pro

115 120 125

Ser Val Leu Lys Phe Val Asp Val Arg Cys Ile Pro Asp Trp Val Ser

130 135 140

Thr Trp Phe Ser Ser Trp Tyr Asn Leu Asn Lys Leu Ile Leu Glu Phe

145 150 155 160

Arg Arg Glu Glu Val Ala Cys Thr Gly Ser Ile Ile Cys Lys Thr Ile

165 170 175

Gly Ser Ile Met Phe Ile Ile Ser Ser Leu Gly Cys Val Ile Lys Ser

180 185 190

Asn Lys Ser Lys Arg Ile Ser Phe Met Thr Tyr Asn Gln Val Leu Thr

195 200 205

Trp Lys Asp Val Met Leu Ser Arg Phe Asn Ala Asn Leu Cys Val Trp

210 215 220

Ile Ser Asn Ser Leu Asn Lys Asn Gln Glu Gly Leu Gly Leu Arg Ser

225 230 235 240

Asn Leu Gln Gly Thr Leu Val Asn Lys Leu Tyr Glu Ile Val Asp Ser

245 250 255

Met Leu Ser Val Cys Ser Asn Glu Gly Phe Thr Leu Val Lys Glu Phe

260 265 270

Glu Gly Phe Ile Met Ser Glu Ile Leu Lys Ile Thr Glu His Ala Gln

275 280 285

Phe Ser Thr Arg Phe Arg Asn Thr Leu Leu Asn Gly Leu Val Asp Gln

290 295 300

Leu Val Lys Met Arg Gly Leu Asn Arg Lys Arg Val Ser Gly Thr Val

305 310 315 320

Leu Glu Gly Asn Gln Tyr Pro Met Tyr Glu Thr Thr Leu Ala Thr Leu

325 330 335

Gly Glu Ala Leu Lys Thr Ile Arg Leu Leu Val Asn Lys Asn Leu Asp

340 345 350

Asn Ala Ala Glu Leu Tyr Tyr Ile Phe Arg Ile Phe Gly His Pro Met

355 360 365

Val Glu Glu Arg Glu Ala Met Asp Ala Val Arg Leu Asn Asn Glu Ile

370 375 380

Thr Lys Ile Leu Lys Leu Glu Ser Leu Thr Glu Leu Arg Gly Ala Phe

385 390 395 400

Ile Leu Arg Ile Ile Lys Gly Phe Val Asp Thr Asn Lys Arg Trp Pro

405 410 415

Lys Ile Lys Asn Leu Lys Val Leu Ser Arg Arg Trp Val Met Tyr Phe

420 425 430

Lys Ala Lys Ser Tyr Pro Ser Gln Leu Glu Leu Ser Ser Gln Asp Phe

435 440 445

Leu Glu Leu Ala Gly Val Gln Phe Glu Gln Glu Phe Ala Ile Pro Glu

450 455 460

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

465 470 475 480

Lys Asn Leu Ile Trp Ser Val Phe Pro Lys Asn Tyr Leu Pro Thr Asn

485 490 495

Ile Arg Glu Lys Phe Thr Asp Glu Met Phe Asn Ser Ser Glu Lys Leu

500 505 510

Lys Thr Arg Arg Val Leu Glu Tyr Tyr Leu Lys Asp Asn Lys Phe Asp

515 520 525

Gln Asn Asp Leu Lys Lys Tyr Val Val Arg Gln Glu Tyr Leu Gly Asp

530 535 540

Lys Glu His Val Val Ser Leu Thr Gly Lys Glu Arg Glu Leu Ser Val

545 550 555 560

Gly Arg Met Phe Ala Met Gln Pro Gly Lys Gln Arg Gln Val Gln Ile

565 570 575

Leu Ala Glu Lys Leu Leu Ala Asp Asn Ile Val Pro Phe Phe Pro Glu

580 585 590

Thr Leu Thr Lys Tyr Gly Asp Leu Glu Leu Gln Arg Ile Met Glu Ile

595 600 605

Lys Ser Glu Leu Ser Ser Val Lys Ser Arg Arg Asn Asp Ser Tyr Asn

610 615 620

Asn Tyr Ile Ala Arg Ala Ser Ile Val Thr Asp Leu Ser Lys Phe Asn

625 630 635 640

Gln Ala Phe Arg Tyr Glu Thr Ser Ser Val Cys Ala Asp Val Val Asp

645 650 655

Glu Leu His Gly Thr Gln Ser Leu Phe Cys Trp Leu His Leu Thr Val

660 665 670

Pro Leu Thr Thr Met Ile Cys Thr Tyr Arg His Ala Pro Pro Glu Thr

675 680 685

Glu Gly Val Tyr Asp Ile Asp Lys Ile Lys Glu Gln Ser Gly Leu Tyr

690 695 700

Arg Phe His Met Gly Gly Ile Glu Gly Trp Cys Gln Lys Leu Trp Thr

705 710 715 720

Met Glu Ala Ile Ser Leu Leu Asp Val Val Ser Val Lys Asn Arg Val

725 730 735

Gln Leu Thr Ser Leu Leu Asn Gly Asp Asn Gln Ser Ile Asp Val Ser

740 745 750

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

755 760 765

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

770 775 780

Ile Gly His Lys Leu Lys Glu Gly Glu Thr Tyr Ile Ser Arg Asp Leu

785 790 795 800

Gln Phe Met Ser Lys Val Ile Gln Ser Glu Gly Val Met His Pro Ser

805 810 815

Pro Ile Lys Lys Ile Leu Arg Val Gly Pro Trp Ile Asn Thr Ile Leu

820 825 830

Asp Asp Ile Lys Thr Ser Ala Glu Ser Ile Gly Ser Leu Cys Gln Glu

835 840 845

Leu Glu Phe Arg Gly Glu Ser Leu Leu Val Ser Leu Ile Leu Arg Asn

850 855 860

Phe Trp Leu Tyr Glu Leu Trp Met His Glu Ser Lys Ser His Pro Leu

865 870 875 880

Ala Gly Lys Gln Leu Tyr Arg Gln Leu Ser Lys Thr Leu Ala Ile Thr

885 890 895

Gln Lys Phe Phe Gly Ile Thr Lys Glu Thr Asp Val Val Asn Leu Trp

900 905 910

Met Asn Val Pro Met Gln Phe Gly Gly Gly Asp Pro Val Val Leu Tyr

915 920 925

Arg Ser Phe Tyr Arg Arg Thr Pro Asp Phe Leu Thr Glu Ala Val Ser

930 935 940

His Met Ser Val Leu Leu Lys Val Tyr Gly Lys Ala Lys Glu Gly Ser

945 950 955 960

Lys Lys Asp Phe Phe Lys Ala Leu Leu Ser Val Asp Lys Asn Lys Arg

965 970 975

Ala Thr Leu Thr Thr Leu Met Arg Asp Pro Gln Ala Val Gly Ser Glu

980 985 990

Arg Gln Ala Arg Val Thr Ser Glu Ile Asn Arg Ala Ala Val Thr Ser

995 1000 1005

Val Leu Ser Leu Ser Pro Asn Gln Leu Phe Cys Asp Ser Ala Ile His

1010 1015 1020

Tyr Ser Arg Asn Glu Glu Glu Val Gly Leu Ile Ala Gln Asn Ile Thr

1025 1030 1035 1040

Pro Val Tyr Pro His Gly Leu Arg Val Leu Tyr Glu Ser Leu Pro Phe

1045 1050 1055

His Lys Ala Glu Lys Val Val Asn Met Ile Ser Gly Thr Lys Ser Ile

1060 1065 1070

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

1075 1080 1085

Arg Ala Val Ser Met Met Leu Glu Asn Leu Gly Leu Leu Ser Arg Ile

1090 1095 1100

Leu Ser Val Ser Gln Asp Asp Ile Ile Leu Pro Thr Lys Ala Asn Gly

1105 1110 1115 1120

Asp Leu Ile Cys Cys Gln Val Ser Arg Thr Leu Arg Glu Arg Ser Trp

1125 1130 1135

Asp Asn Met Glu Ile Val Gly Val Thr Ser Pro Ser Ile Val Thr Cys

1140 1145 1150

Met Asn Ile Val Tyr Ser Ser Ser Ser Gln Leu Lys Gly Ile Thr Ile

1155 1160 1165

Glu Lys Phe Ser Thr Asp Lys Thr Thr Arg Gly Gln Arg Gly Pro Lys

1170 1175 1180

Ser Pro Trp Val Gly Ser Ser Thr Gln Glu Lys Lys Leu Val Pro Val

1185 1190 1195 1200

Tyr Asn Arg Gln Ile Leu Ser Lys Gln Gln Lys Glu Gln Leu Glu Ala

1205 1210 1215

Ile Gly Lys Leu Arg Trp Val Tyr Lys Gly Thr Gln Gly Leu Arg Arg

1220 1225 1230

Leu Leu Asp Lys Ile Cys Ile Gly Ser Leu Gly Ile Ser Tyr Lys Asn

1235 1240 1245

Val Lys Pro Leu Leu Pro Arg Phe Met Ser Val Asn Phe Leu His Arg

1250 1255 1260

Leu Ser Val Ser Ser Arg Pro Met Glu Phe Pro Ala Ser Val Pro Ala

1265 1270 1275 1280

Tyr Arg Thr Thr Asn Tyr His Phe Asp Thr Ser Pro Val Asn Gln Thr

1285 1290 1295

Leu Ser Glu Arg Phe Gly Asn Glu Asp Ile Asn Leu Val Phe Gln Asn

1300 1305 1310

Ala Ile Ser Cys Gly Ile Ser Val Met Ser Val Val Glu Gln Leu Thr

1315 1320 1325

Gly Arg Ser Pro Lys Gln Leu Val Met Ile Pro Gln Leu Glu Glu Ile

1330 1335 1340

Asp Ile Met Pro Pro Pro Val Phe Ser Gly Lys Phe Asp Tyr Lys Leu

1345 1350 1355 1360

Val Glu Lys Ile Ser Ser Asp Gln His Ile Phe Ser Pro Asp Lys Leu

1365 1370 1375

Asp Leu Val Thr Leu Gly Lys Met Leu Met Pro Ser Thr Ser Gly Ala

1380 1385 1390

Lys Ser Asp Gln Phe Trp Asn Arg Lys Glu Asn Phe Phe His Gly Asn

1395 1400 1405

Asn Leu Val Glu Ser Leu Ser Ala Ala Leu Ala Cys His Trp Cys Gly

1410 1415 1420

Ile Leu Thr Glu Gln Cys Asn Glu Asn Asn Ile Phe Arg Arg Glu Trp

1425 1430 1435 1440

Gly Asp Gly Phe Val Thr Asp His Ala Phe Ile Asp Phe Lys Ile Phe

1445 1450 1455

Ile Gly Val Phe Lys Thr Lys Leu Leu Cys Gly Trp Gly Ser Arg Gly

1460 1465 1470

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

1475 1480 1485

Ile Arg Val Asp Asn Ser Phe Trp Arg Met Phe Ser Lys Val Ile Leu

1490 1495 1500

Glu Pro Lys Val Arg Lys Arg Val Met Leu Phe Asp Val Lys Ile Leu

1505 1510 1515 1520

Ser Leu Val Gly Tyr Ala Gly Phe Lys Asn Trp Phe Ile Asp His Leu

1525 1530 1535

Arg Ser Ser Asp Leu Cys Glu Val Pro Trp Val Val Asn Ala Asp Ser

1540 1545 1550

Glu Ile Val Glu Val Ser Ala Val Lys Ile Tyr Leu Gln Leu Leu Arg

1555 1560 1565

Val Ser Ser Pro Leu Arg Ile Thr Val Leu Asn Tyr Ser Asp Met Ala

1570 1575 1580

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

1585 1590 1595 1600

Pro Ser Ile Ser Arg Thr Leu Ser Pro Ala Glu Leu Ala Pro Val Val

1605 1610 1615

Glu Pro Thr Val Gln Met Asn Leu Phe Pro Lys Ile Thr Phe Glu Arg

1620 1625 1630

Leu Lys Asn Tyr Glu Thr Ile Ser Gly Ser Thr Arg Gly Lys Leu Thr

1635 1640 1645

Arg Asn Tyr Met Val Met Leu Pro Trp Gln His Ile Asn Arg Phe Asn

1650 1655 1660

Phe Val Phe Ser Ser Thr Gly Cys Lys Ile Ser Val Lys Ser Cys Ile

1665 1670 1675 1680

Gly Lys Leu Ile Gln Asp Leu Asn Pro Thr Val Phe Tyr Phe Val Gly

1685 1690 1695

Glu Gly Ala Gly Asn Trp Met Ala Arg Thr Ala Cys Glu Tyr Pro Asn

1700 1705 1710

Thr Lys Phe Val Tyr Arg Ser Leu Lys Asp Asp Leu Asp His His Phe

1715 1720 1725

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

1730 1735 1740

Gly Gly Glu Gly Leu Ser Met Asp Thr Thr Asp Ala Thr Gln Lys Thr

1745 1750 1755 1760

His Trp Asp Leu Thr His Arg Ile Cys Lys Asp Ala Leu Leu Ile Thr

1765 1770 1775

Leu Cys Asp Ala Glu Phe Lys Asp Arg Asp Asp Phe Phe Lys Met Val

1780 1785 1790

Thr Leu Trp Arg Lys His Val Leu Ser Cys Arg Ile Cys Thr Thr Tyr

1795 1800 1805

Gly Thr Asp Leu Tyr Leu Phe Ala Lys Tyr His Ala Lys Glu Glu Ser

1810 1815 1820

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

1825 1830 1835 1840

Gly Ser Lys Leu Ser Gly Ser Glu Cys Tyr Val Leu Leu Thr Leu Gly

1845 1850 1855

His His Asn Asn Leu Pro Cys His Gly Glu Val Gln Ser Ser Lys Leu

1860 1865 1870

Lys Met Ala Val Cys Asn Asp Phe Ser Ile Pro Arg Lys Val Glu Val

1875 1880 1885

Lys Ala Val Glu Ala Asn Cys Lys Ser Leu Leu Ser Gly Leu Arg Thr

1890 1895 1900

Pro Ile Asn Arg Ala Glu Leu Asp Arg Gln Lys Lys Met Leu Thr Leu

1905 1910 1915 1920

Arg Ser Tyr His Ser Ser Val Ala Thr Val Gly Gly Ser Arg Val Ile

1925 1930 1935

Glu Ser Lys Trp Leu Ser Lys Lys Ala Thr Thr Ile Ile Glu Trp Leu

1940 1945 1950

Glu His Ile Leu Asn Ser Pro Lys Gly Glu Leu Asn Tyr Asp Phe Phe

1955 1960 1965

Glu Ala Leu Glu Asn Thr Tyr Pro Asn Met Val Lys Leu Leu Asp Asn

1970 1975 1980

Leu Gly Ser Ala Glu Leu Lys Lys Leu Ile Lys Val Thr Gly Tyr Met

1985 1990 1995 2000

Leu Met Ser Lys Lys

2005

Claims

1.一种鸭C亚型禽偏肺病毒弱毒株，保藏号为：CCTCC NO:V201823；核苷酸序列为SEQID NO：1。

2.根据权利要求1所述的番鸭C亚型禽偏肺病毒弱毒株，其特征在于，所述番鸭C亚型禽偏肺病毒弱毒株的阅读框氨基酸序列为SEQ ID NO：2至SEQ ID NO：9。

3.一种番鸭C亚型禽偏肺病毒弱毒株的制备方法，其特征在于，包括：将番鸭C亚型禽偏肺病毒S-01株进行传代培养至40-62代。

4.根据权利要求3所述的番鸭C亚型禽偏肺病毒弱毒株得制备方法，其特征在于，所述番鸭C亚型禽偏肺病毒弱毒株的制备方法，包括，将C亚型禽偏肺病毒S-01株进行传代培养至50代。

5.根据权利要求3所述的番鸭C亚型禽偏肺病毒弱毒株得制备方法，其特征在于，所述番鸭C亚型禽偏肺病毒弱毒株的制备方法，具体包括：

6.权利要求1-2任意一项所述的番鸭C亚型禽偏肺病毒弱毒株或权利要求3-5任意一项所述的方法所制备的弱毒株在制备番鸭免疫产品中的应用。

7.一种包含权利要求1-2任意一项所述的番鸭C亚型禽偏肺病毒弱毒株或权利要求3-5任意一项所述的方法所制备的弱毒株的番鸭免疫用制品，其特征在于，每份免疫制品中所述弱毒株的含量为≧10^2.5TCID₅₀。

8.根据权利要求7所述的番鸭免疫用制品，其特征在于，每份免疫制品中所述弱毒株的含量为≧10^4.5TCID₅₀。