专利名称：日本血吸虫雌虫性别特异基因、编码蛋白及其应用的制作方法
技术领域：
本发明涉及生物基因领域，尤其涉及日本血吸虫雌性成虫特异蛋白质的基因核酸序列、以及特异蛋白质的氨基酸序列；此外，本发明还涉及日本血吸虫雌性成虫特异基因、蛋白的应用。
背景技术：
血吸虫病由血吸虫的感染引起，在世界上有76个国家，受感染者约1.5亿。致病的血吸虫有3种，既日本血吸虫(Schistosoma japonicum)，曼氏血吸虫(S.mansoni)和埃及血吸虫(S.haematobium)。在我国流行的是日本血吸虫。血吸虫病的流行给人民身体健康和经济发展都造成巨大的损失，因此，防治血吸虫病、开发研制治疗血吸虫病和抗血吸虫的药物具有巨大的社会和经济效益。
研制新的治疗血吸虫病和抗血吸虫的药物，首先必须找到合适的药物作用靶点。血吸虫的虫卵是造成血吸虫病的最主要原因，肉芽肿就是由虫卵造成的。此外，虫卵排出体外进入外界环境，污染环境后会制造更多的感染。所以，治疗血吸虫病、防止血吸虫的扩散，一个关键的措施是抑制血吸虫雌虫成虫的产卵。这可以通过干扰雌虫性成熟，干扰雌、雄虫的配对，或者抑制产卵相关的生理途径来实现。而雌虫特异表达的蛋白质，很可能与雌虫性成熟、雌雄虫的配对，或者产卵密切相关。通过大规模的基因表达谱学研究，发现了日本血吸虫雌性成虫的特异蛋白质，这些蛋白质，都是可能的药物作用靶点和候选疫苗。
在本发明之前，还没有出现涉及本发明的日本血吸虫雌性成虫特异蛋白的公开报道。
发明内容
本发明要解决的技术问题之一是提供一类日本血吸虫雌性成虫特异蛋白，及其基因的核酸序列。
本发明要解决的技术问题之二是提供一类日本血吸虫病防治疫苗，该疫苗以日本血吸虫雌性成虫特异蛋白为免疫原。
本发明要解决的技术问题之三是提供一类与日本血吸虫雌性成虫特异蛋白结合的抗体。
本发明要解决的技术问题之四是提供日本血吸虫雌性成虫特异蛋白在制备预防和治疗日本血吸虫病的药物中的应用。
为了解决上述技术问题，本发明通过如下技术方案实现在本发明的一个方面，提供了一类日本血吸虫雌性成虫特异蛋白，包括(a)具有SEQ ID No.25～SEQ ID No.48所示氨基酸序列的蛋白；(b)由(a)的蛋白发生一个或几个氨基酸置换、缺失或插入而形成的具有(a)蛋白相同活性的蛋白。
较佳地，所述蛋白是具有SEQID No.25～SEQ ID No.48氨基酸序列的蛋白。
在本发明的另一方面，提供了一类编码日本血吸虫雌性成虫特异蛋白的基因，包括(a)SEQ ID No.1～SEQ ID No.24所示的核苷酸序列；(b)SEQ ID No.1～SEQ ID No.24所示的核苷酸序列中每条序列的互补序列；(c)与SEQ ID No.1～SEQ ID No.24所示的核苷酸序列中每条序列有至少70％同源性、且编码相同功能蛋白质的核苷酸序列。
较佳地，所述基因具有SEQ ID No.1～SEQ ID No.24所示的核苷酸序列。
在本发明的另一方面，还提供了一类日本血吸虫病防治疫苗，该疫苗以日本血吸虫雌性成虫特异蛋白为免疫原。
在本发明的另一方面，还提供了一类与日本血吸虫雌性成虫特异蛋白结合的抗体。
此外，本发明还提供了日本血吸虫雌性成虫特异蛋白在制备预防和治疗日本血吸虫病的药物中的应用。
在本发明中，术语“日本血吸虫雌性成虫特异蛋白”指具有日本血吸虫雌性成虫特异蛋白活性的SEQ ID No.25～SEQ ID No.48所示氨基酸序列的蛋白质。该术语还包括具有与日本血吸虫雌性成虫特异蛋白质相同功能的、SEQ ID No.25～SEQ IDNo.48序列的变异形式。这些变异形式包括(但并不限于)若干个(通常为1-50个，较佳地1-30个，更佳地1-20个，最佳地1-10个)氨基酸的缺失、插入和/或取代，以及在C末端和/或N末端添加一个或数个(通常为20个以内，较佳地为10个以内，更佳地为5个以内)氨基酸。例如，在本领域中，用性能相近或相似的氨基酸进行取代时，通常不会改变蛋白质的功能。又比如，在C末端和/或N末端添加一个或数个氨基酸通常也不会改变蛋白质的功能。该术语还包括日本血吸虫雌性成虫特异蛋白的活性片段和活性衍生物。
在本发明中，术语“编码日本血吸虫雌性成虫特异蛋白的基因序列”指编码具有日本血吸虫雌性成虫特异蛋白活性的核苷酸序列，如SEQ ID No.1～SEQ ID No.24所示的核苷酸序列及其简并序列。该简并序列是指，位于SEQ ID No.1～SEQ ID No.24序列中，有一个或多个密码子被编码相同氨基酸的简并密码子所取代后而产生的序列。由于密码子的简并性，所以与SEQ ID No.1～SEQ ID No.24所示核苷酸序列的同源性低至约70％的简并序列也能编码出SEQ ID No.25～SEQ ID No.48所示的氨基酸序列。
在本发明中，可以使用一系列本领域已知的方法来制备针对日本血吸虫雌性成虫特异蛋白的抗体。例如，将提纯的日本血吸虫雌性成虫特异蛋白产物或它的抗原片段注射入动物体内以产生多克隆抗体。同样，表达日本血吸虫雌性成虫特异蛋白或它的抗原片段的细胞也可以用来对动物致免疫而产生抗体。本发明制备的抗体也可以是单克隆抗体，这些单克隆抗体可用杂交瘤技术制备。
应用本发明的日本血吸虫雌性成虫特异蛋白，可抑制血吸虫雌性成虫的产卵，从而治疗血吸虫病、防止血吸虫的扩散，该类蛋白在血吸虫病防治和治疗中有着重要的作用。
图1是用RT-PCR实验验证日本血吸虫成虫特异表达基因的图；图2是图1的数据分析图。
具体实施例方式
下面结合附图和实施例对本发明作进一步详细的说明。
以下实施例仅用于说明本发明而不用于限制本发明的范围。实施例中未注明具体条件的实验方法，通常按照常规条件，例如Sambrook等人，分子克隆实验室手册(New YorkCold Spring Harbor Laboratory Press，1989)中所述的条件，或按照制造厂商所建议的条件。
实施例1日本血吸虫雌性成虫特异蛋白的生物信息学分析。
1.日本血吸虫雌性成虫特异蛋白的筛选利用表达序列标签(EST)的数据，分析特异表达的蛋白。雌虫相对于雄虫特异表达的基因1)其差异显著性必须满足p＜0.05，这个显著性分析用网上专门的软件工具计算获得；2)每个基因，其EST数量必须大于等于10条；3)雌虫的EST数量必须是雄虫数量的3倍或3倍以上。经过这3个标准的筛选，得到了雌性成虫特异表达基因。
2.同源性和相似性分析日本血吸虫雌性成虫特异蛋白序列对GenBank(http://www.ncbi.nlm.nih.gov/)的非冗余数据库(Non-redundant，NR)做BLASTP比对；日本血吸虫雌性成虫特异蛋白序列对Swissprot数据库做BLASTP同源性比对。根据比对结果得到的与已知其他物种或已知的血吸虫蛋白的序列相似性，做出日本血吸虫雌性成虫特异蛋白的功能注释。
实施例2RT-PCR实验验证日本血吸虫雌性成虫特异蛋白的表达情况。
1.样品准备准备日本血吸虫的成年雌虫和雄虫，以抽提其RNA。
2.总RNA抽提试剂盒抽提RNA试剂采用TRIzol reagent(GIBCO/BRL)，该试剂基于酸性酚一步抽提法生产。用于抽提RNA所用器皿和水均要进行无RNA酶处理，以保证实验中无RNA酶环境。
3.RNA抽提步骤将碾杵和匀浆器等器皿在200℃干烤4h，去除RNA酶，冷却；加入液氮中预冷，将组织从液氮中迅速取出，碾成粉末；用刮匙将组织放入预先加入TRIzol试剂匀浆器中，匀浆数分钟；将匀浆后液体转入无RNA酶离管中，加入氯仿后，4℃离分层；将上层水相转入一无RNA酶离管中，加入氯仿后，4℃离分层；将上层水相转入一无RNA酶离管中，加异丙醇，4℃离沉淀RNA；用75％乙醇洗涤沉淀2次；用无RNA酶去离子水溶解沉淀。抽提RNA质量鉴定紫外分光光度计测定260/20比值(比值均在1.7~2.0)；并在MOPS甲醛变性胶中观察有无降解。
4.RT-PCR反应用半定量反转录多聚酶链式反应(RT-PCR)来验证日本血吸虫雌性成虫中一些基因的表达丰度差异。采用不同的循环数，分别是21、24、27和30个反应循环。每个PCR重复3次。实验结果见图1和图2所示。
序 列 表<110>上海人类基因组研究中心<120>日本血吸虫雌虫性别特异基因、编码蛋白及其应用<130>NP-10266<160>48<170>PatentIn version3.3<210>1<211>1442<212>DNA<213>日本血吸虫(Schistosoma japonicum)<400>1gttttgcgga gcgttgttaa ttagtgatgc gtgaatgtat cagtcttcat gttggtcagg60ccggagccca aatgggcgac tcttgttggg agctctactg ccttgagcat ggtatacaac 120ctgatggaaa ggttcctagc gactctacta taaaaaatga caacgattct ttcagtacat 180tttttagcga gaccagctat ggccgttacg tcccacgcgc tgtgttcgtc gacttagaac 240cgtcggttat tgacagactg cgaacagggg cttataagca gttattccat cctgaacaaa 300tgataagtgg aaaagaagat gctgccaata attacgctcg cggtcactat actgttggta 360aggagttggt tgatgaagtc cttgaccgca tacgcaagtt gtctgaaaga tgcacagggc 420ttcagggatt cttggtattt cattcatttg gcggtggaac aggatcagga ttcacgtctt 480tgctgatgga acgcctctcg gttgatttcg gtaaaaaagc caagttagag tttgctgttt 540acccagcacc tcatatctcg acagctgttg ttgaacccta taactccatt ctaaccacgc 600atacaacact agaacattct gattgcgcgt ttatggtcga taatgaagcc atttatgaga 660tctgtcgccg taatttaact atcgatcgac caacatattg caaccttaac cgattaattg 720gacagatcgt cagttcaatt actgcctcac tacgatttag tggtgcttta aacgttgatt 780tgaacgaatt tcaaactaat ttggtccctt atccacgtat ccacttccca ctaacaacct 840atgctcccat tatttcagca gagaaagctt tccatgaaca actgagtgtt tcagaaataa 900ctaactcctg ctttgaacct gcaaatcaga tggtcaaatg tgacccaaga aatggaaaat 960atatggcatg ttgtttgtta taccgtggtg acgttgtgcc taaggatgtg aatgccgcaa 1020tcgcgaatat aaagactaga cggtccatcc agtttgtcga ttggtgtcct acaggtttca 1080aagttggaat caactaccaa cctccaactg ttgttccagg tggagatcta gcaaaggttc 1140agcgtgctgt ttgtatgctt agtaatacga cggcgatatc ggaagcttgg gcccgtttaa 1200atcggaagtt tgatctaatg tatgctaaac gtgcatttgt tcattggtac gttggtgaag 1260gcatggagga aggagaattt tcagaagctc gcgaagactt ggctgctctt gaaaaagatt 1320acgaagaagt tgcttcagat actgtagatg cacagtctga tgatggattt taatgtaata 1380tgattacaat ttacaaaata aatttttata ttctgtattt ctctaaaaaa aaaaaaaaaa 1440aa 1442
<210>2<211>1276<212>DNA<213>日本血吸虫(Schistosoma japonicum)<400>2tgtggttatc ctatcccatg atgcctgcct atactaatgg tggaaatgaa gagttggaaa60ccttcctgtt cacttctgaa tctgttggtg aaggacatcc agataaactt tgtgatcaga 120taagtgatgc catacttgat gagtttttga agcaggatcc cgatgccaaa gtagcatgtg 180aatcgtttgt caaaagcggt cttattattg ttgggggcga aatttcttct aatgctgtga 240tcgacattca gaagaccgtc cgtaatgtcg ttaaagacat tggctatgat gatactcgca 300aaggtcttga ctataaaaat tgcaacatct tagctgtgat tgaacaacag agtgcagata 360ttgcgaaagg tgtgcatgaa ggaagaaatg ttgaagacat tggtgctgga gaccaaggtt 420taatgttcgg ctatgcgact gacgaaactg aagaatgtat gccattaacg gttaccttat 480cacatgcttt aaataaacgc ctggctatgt gcagacgtaa tggcattttg ccttgggccg 540gtcctgattc aaaaactcaa gtgactgttg agtatttaaa tgaccatgga gcatgtcgac 600ctttacgagt acatacagta gtcatatctg cacaacacga cgagcatgta accttagaga 660aagttcgtgc tgatttaatg gaatacgtta ttaaagagga aatacctgct aaccttttgg 720atgaaaaaac aaaatatcat ttgaatccag ctgggagatt tgtaattggt ggtcctcaaa 780gcgatgctgg tttgacagga agaaaaatca ttgtagatac atatggaggc tggggtgcac 840atggtggcgg agcgttttct gggaaggatt actcaaaagt agaccgatct gcagcttatg 900cagcacgatg ggtgtctggt ctttgtagac gtgtcttggt tcagctttct tacgctatcg 960ggattgctga accattgtct attaacgtaa acagttatgg aactggcaag attagtgata 1020agaagctact ggatgttatt atgaagaatt ttgatttgcg acctggtata atagtcaagg 1080atctagacct tcgaagacca cgttacttac aaacctccgt ctatggtcac ttcggtcgac 1140ctgaatttcc ttgggaacaa tgtagaaagt taaaattgta atattgacga ttttgtccag 1200ttccatttct attttagaaa tatctattca tatgaatata aataataaat tggaaattta 1260aaaaaaaaaa aaaaaa 1276<210>3<211>1492<212>DNA<213>日本血吸虫(Schistosoma japonicum)<400>3gattcatgaa cttttaaagt gacagctagt taccccacac ttttattgcc cgggtttatt60cagatcaatt acttccatgg ctgaaataaa gtatcgtagg tgtttgtcgc tttcaatcta 120ttgttagtgg cgaagtactg tcgatatttt cagtatggag cagtacattt catacagtct 180atgaaaacct aacgtaatca gtactatacg ttcctttcta gaacacctgc tctcattcag 240ggatttttta ttttacatca cttcttttct agtcccgaat ttcagtaata gtaaacaatt 300tatgttatga aatccatttt gcaagcatag atattgaatt ctgtgaattt ggttgtttct 360catacaaatt tacttcaata tcggttatat ttagtcaaaa cattttagaa attgttaacc 420taactattca ctatcacttc atatgaatca tacaagagca ttacattttg ttttcaaagt 480agctaatcgt aatgaaacaa tagattttta caaacgaatt cttggaatga aggttttaag 540
acatgaagaa ttcagtgagg gttgtaaagc ttcctgcaat ggaccatatg atggaaagtg 600gagtaaaact atgatagggt atggtccgga agataatcat tttgttattg agttgactta 660taattatggg gtcggtgctt atcaacttgg taatgatttc caatatatac gtattcacaa 720caaggaagcg tattctagaa taatcaatgg atcttggcca atcacgtcag aagaatcaga 780ttcagtaaag gtgaaagctc ccggcggata tacattttgg gtgcataact ctgatagcga 840tggggatcca gttcgaatgg tggctttaag tagttccgat attaaacgat caattgatta 900ttggtcacac aaacttcata tgacacttgt atcatccagt tcaaatgaag ccgtatttta 960ctattcaccg aatcaatgta atttgaagtt aatttctatt caaaagcctg ttgatcatgc 1020tagtgctttt ggtcgtatag cttttgcatg cccaagtgct caattaccta ttttgcaaga 1080attaatggac aaagaaaatg agaccattct tacgcgttta gtcagtctgg acacaccagg 1140aaaggctaca gtacaagtta ttattctagc tgatccagat ggtcatgaaa tttgctttgt 1200tggagatgaa gcatttagac aattatcaca agtagattca caagctgata gtctacttca 1260aactgctata acttcagata aaagtgatga atggtacaat aaacatggtg gaaagacaac 1320aaaatgaaat gaaatacaat ttcatttgaa ctcattcaat taagtatact aatggtggga 1380taatttagtc actatttttt gtaccttttt ctttttcata cggaatattt gattttcttt 1440gctttaatga agaaattatt gttgattctc ttacaataag tcattagata gt 1492<210>4<211>757<212>DNA<213>日本血吸虫(Schistosoma japonicum)<400>4taaatatatg tatatatgtg tgttcgagta tggagaaaaa atatgtagtt atccaataac 60aaggtattga ggataaagat gaagttgcat tgattgcatt ttgttacggt tatgtttaag 120agaaacacat tattgatcag tttataatga aattgtgtaa tagtctatca tgttgtagga 180gtttttggct tggcgaaacc caaattactt tgattaaata ccacacttcc tagattacca 240tagctttcaa tgataatttc accattagta aactgaacat tcataccaga acaagcttct 300ttcaaagctg caagaaaatt gttcacatca tatacagata ctggattaga taaatcttcc 360agccgctagc atcaaaccgc tcgaaaaagt cgctgatggt ctcattaaga aagctaaaga 420ccagaatttg agggttaaag gccctgttcg aatgccgaca aaagttttga gggttactac 480gcggaagact ccatgtggtg aaggctcaaa gacatgggac aggtttcagt taagaatcca 540caagcgtgtg atcgatttac tgtctactga cgaagcgaca ctacagttga aacacatcta 600tttagagcct ggcgtcaact gtgatgttaa agtttcagga gatttatagc ttgtaatata 660attatgtgaa aggatctcta gacttttgtt aaagtgcttt gttacatgta gaacttaccg 720tcgtagtgaa tccattcaat aaaatattca ccctatt 757<210>5<211>1756<212>DNA<213>日本血吸虫(Schistosoma japonicum)<400>5ttgacttatt agacttatca aaccctgacg ctaaaccttc aaattcttcg aacaatatag 60
ttcagaagcc tactcatgtt actgaaaatg gcctaaaata caatcatgat agcaatctta120cgcattcggt gtcacctcct aaagaacaac tattattaca gttagatgtt actcaagctc180gtcttgttag gaattatggt ttcctcccta tgaaccttta ctgcataatt cgcattggag240atgccagatt cgaaactcag acttcatcaa gatctggtaa acatcccatc tggaatgaaa300cttttcgatg ttggctccat gatggtgttg acaaattatc attagaaata gtttcggaaa360atatattttc gtcttcacaa gtcgctcatg ctgtaatagc ttttccacga tcgttatatg420atggtatcgc tttaaatcag tggtttgagc ttagtggaca acaaggagaa aacttagaag480gttcaattaa tctgattttc caatgtcgaa aagtaccagt aaatctccca cttctccaat540cgactccact atactgtcaa ccaagattgg gtgtagttgg tgatatgcaa gttttaccta600aaccgatatc tactgaagaa gttcagtcaa tacatgaaat cttcccatct gtcgaagagg660atacaattcg atcactttta gcaacacatc atggaaatcg tgaaattgta atcagcgaat720tacttctatg gcatcggaaa cttgaaatag tatgttcaga ctgtgatgta tttaaatcat780tttcaaataa tgttattttt ctgttcattt gttattggaa cgatttttct tctgaacaac840gttttccgct cacagcacta acgtgttatt accctgtttt acttcaacct cctttatcaa900actttttaac aaaaataaaa cagcgttatg atcatttcta cactgatttt atctaattca960attatcagtt attatttgtt tgttttcaca cagttacagt atatgtgcat tctgtgcgta 1020tcttcatcct tctttcctaa tacctaatta ttaagatcat ctcaatgaaa cgacactgta 1080gttataatgt ttatcacgta tgtatatttt attttgttgt ccatgttgtt cttattttcc 1140attttttcta ccaatttatt ttcacctcta agcatatcct atttttgtgc tacatcagtt 1200tgttatatag ccaaaacttt agtgaattca atgatagtgt ctatctcttt acaactgtca 1260ctactgtcgt tgtaactgtc actcttttgt cttctacaaa taatgattgt gtagtttata 1320tatatatata tatatatata tatatatata tatatatata tatatatttc gaatcatcaa 1380cctatggatt gaaactaact tctctaacta ccgtttcatt tacttaccaa atatagattt 1440ttgtatactc tgctttaagt aacattgttt gtgtacgaac aattgtctta aatatactag 1500attgaataag gttagtaact tcaaacccat tgatcgaaaa tgaatacttt ttcaaatact 1560tggacactac tctccattta tgtatagtag gcttatggaa ttgctagtca tgtccccact 1620ttgtatcctg cctagagcct ttaggctcca cggtaaatgc ttaaccctct gtccattgaa 1680cctatatcca gtgattgaca actcacagta tatttctcag tcttgtccct gccattggtg 1740agtaccgatc acctat 1756<210>6<211>1760<212>DNA<213>日本血吸虫(Schistosoma japonicum)<400>6tttgtaaacc acaatacggc tgacatcaga gcccatagtt tcaaagagct ccacctcaaa 60cttaagagat ttcccatcca tgtacgcata ttcaggcgac gcaatcaatt cacaaacttc120tcccaacctc atagttgcaa caccgatatc ccaagcttta attacactac ctttcccaat180tgtaaattca aacttttcat ttcgggctct gcttgagtcg aatacttcac catgcttttc240accaccgaag ttagtaccga cataatgtac tatgaccgtg tcatcatcgc atggtttgat300gtccgagtag ccttctcgta ctactttctg gaaacattag tggaatgaac aaggacgcag360ttgacatgtc ggagacgcca aagcagtctg aagaggaata tctaaaagat ttcatagact420taagcccatc tggtgatcgt ggtattctca agaaagtagt acgagaaggc tactcggaca480
tcaaaccatg cgatggtgac acggtcatag tacattatgt cggtactaac ttcggtggtg 540aaaagcatgg tgaagtattc gactcaagca gagcccgaaa tgaaaagttt gaatttacaa 600ttgggaaagg tagtgtaatt aaagcttggg atatcggtgt tgcaactatg aggttgggag 660aagtttgtga attgatcgcg tcgcctgaat atgcgtacat ggatgggaaa tctcttaagt 720ttgaggtgga gctttttgaa actatgggct ctgatgtcag cagaaacaaa gatgggagta 780ttcgcaagtc aattattaaa aagggaagag atatccacaa tcctgtagct ggggctgaag 840ccactattgt tttccgcaat cttttgaact tgacggagga aacagaagtc acatattgtg 900ttggcgaccc tccatcaaac gtaccggatg agttggacca atctgttcgt cacatgaata 960caggtgaagt cagccgtatt gtggtttaca aagacggtca ttcattaact tctggagatt 1020cggataaaga tagaatagtt tacgaactaa cactgaagtc ttttgaaaag accaaacacc 1080tctctggtat tacgtctttt ccagagcgaa tagcctatgc aaatacactt aaagagaagg 1140ccaacaattt cttaaaggat tctaaatttg attcagctat cgaattatat aaacgtttgg 1200acgatgagct gcagtacgta gtggctaatg ggcctacaga acaaaaggaa ttgtctgggg 1260tcactgtagc tgtccaactc aatttagctc tggtctatct gaagctttgt aaaccagaca 1320aatgtattga gttttgcaag aaagtgctcg ataattttag cgacaacgaa aaggcgctat 1380ttagaattgg tcaggcacat ttactacgta aggaccatga agaagcagtt gtttacttca 1440aaaggattgt aacaaaaaat cctaacaatg catctgctgt aaaacaggtt caaatctgtg 1500aggaagagat tagaagagca aaagatatag aacgaaagaa gtttcgtcat attttcgaac 1560gttgtaaaga cacgggatta aatgatgctc aaaatcatga ggatggggtt gtagtgaatg 1620atgaaaagtc tgccctgtga gagcttaatt ctgtgtgtgg catgaagtta gcaccccgtg 1680attttgtaaa tgtttcttat aattactggt ttaatttcaa agttgtgaat acattttaaa 1740gcaaacaaaa aaaaaaaaaa 1760<210>7<211>1758<212>DNA<213>日本血吸虫(Schistosoma japonicum)<400>7ttcgttttca tctcgtcatg ctgacaagtg gtgatagtga tctggtgggg gaagctgtta 60agtttcacaa accaggggag aactacaatt ctcctggtta tgttgtgaca aaaaatacgt 120tgcggttatt aaaggagcac ctgaaacaaa cgggtggtaa ggtagtcacc aggtttcccc 180ctgaaccaaa tggtatttta catattggtc atgcaaaggc aatcaatttt aattttggct 240atgcaatgaa gaatggtggt ataacttact tgcgatacga tgatactaac cctgaaaaag 300aagaagcaga attttttgaa gctattgagg atatggtcag gtggttaggt tttacaccat 360ataaaattac gtacgcatcg gataattttc atcagttata cgaatggtct gtagaagaaa 420taagaggttt taatccacct ccttctccat ggagagaaag gccggttgaa gaatctcttc 480gactgtttga agacatgaaa aatggaaaga tcgatgaggg aatggcaact ttacgtttga 540aagtgacgtt gggagatgga aaagtggatc cagtagctta tcgtatcaaa atgactcccc 600accaccgtac tggtaatgaa tggtgtattt atcctactta tgattatact cattgtttat 660gtgattccat cgagaatatt acacattcat tatgcaccaa agagttccag tcacggcgtc 720cagcctacta ttggctttgt aatgcgctag atgtttactg tccagttcaa tgggaatatg 780gtcgtctcaa tttgtattat agtgtagtat ctaagcgcaa aattttaaag ctgatagaag 840ctggcattgt tagtgattgg gatgatccac ggttagtaac actgacagca cttcgtcgtc 900
gtggatttcc gcctcaagct attaatatgt tttgtgaacg tattggcgtt actatggccc 960aaacaatcct cgatccttct gctctagacg cttgcgtacg cgactatttg aatgatcatg 1020ctccacgtgt aatggctgtg ttagagccag tgttggtaac tattactaat tggtttgaat 1080tatacggcga taagtcatct atcgagctta cagtagcgga ttttcctgcc gttccggata 1140gtaaaactca cagcattact ttgcaacatg atctatacat agagaagacg gattttcaag 1200aagttgctga gaaaggatat cgtcgtttaa cgcccactca ggcagttggt cttcgtcaca 1260caggccttgt tttagaattt tctgatttta aaaaggattc taatggggaa atagttcatt 1320tattcgtaaa agccaaaaag gttgaagatt ctattaaacc aaaagctttt atacactggg 1380tatctaatcc gttgcagttt gaagctcgac tgtatgaccg attgttcact gtcaaggaac 1440cagaaagcgc aaaggaggga ttcttgtctg tcattaataa agattcatta gtcgtactct 1500cggatgcttt gattgagcag tctgtcatga atgcaaaagt ttacaccgct tatcagtttg 1560aacgtattgg tttcttttct gttgatccag atacaacctc gaaacgcatg gtcttcaata 1620ggactgtagc tctcaaagtc gatcctggta aaacaatttg atgtgaaaag catctcagtt 1680atcataaatc attaatgacg tgttgaacac tatgtgtaaa gtgtcatgta atttgtaata 1740aatttcttac tctggatc 1758<210>8<211>932<212>DNA<213>日本血吸虫(Schistosoma japonicum)<400>8cgatatttcc gtcgcattaa tacgggatca gaccacgtgt tttagtgtgc gacaatgttt 60taatacttaa acgttatggt gcaccagtga ttttgcgggg ctggatggaa gagttaacta 120cttttgttat acggaggcga gcatattggc cgtgtagaat aataccgatc tggcgacgtg 180cactccacat agatagcgat accaaagacg aggggtcacc cctcacagtg gtgaacgact 240cgtgaaatcg aagtggtctc cccgggcgcg cgctaatttg ggccgccacg atgcgagtgt 300tcaaggatgc tatttctggt gatgagatgt tttcagactc tcactctcct cagcttataa 360acgatgtcgt gtatgaagtc gatgccaact ttataacagt gtctaatggt cttgatagca 420aattgatcgc tgctaatcca tcaggtgaag aaggccaaga agaagtatct gattccactg 480aaagagttat tgatctagtt catgcgagcc gactagtcag tacgtctttc gataagaaat 540catatagagc atatctcaag ggctacttga aagccataaa ggaacgttta caaaaagaga 600atccagaaag agtcagtata tttgaaagtc gaatcaacga atacatggta aatgtcttca 660aaaatttcga cgattatgag cactacattg gcgagtcaat gaaccctgat ggtatggtag 720cactgatgaa ctttcgtgag aatggtgtga ctccctactt tgttttccta aaagatgggt 780tgattgaaga aaaatactaa gtcgcttcag gtcattatcg gaattccgaa ttcatggaaa 840acatcttgta aatcagtttc aatatacaat attttctgga taaaaaaaaa aaaaaaaaaa 900aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aa932<210>9<211>951<212>DNA<213>日本血吸虫(Schistosoma japonicum)
<400>9atcatttaga atggataaga tttatttatc tggaattcac acattacgat attacagtga 60gatgaattga gtggtgtgtg ttgtatgatg agaggaatga gagcagtatg acacaaatga120atgttggagt gatatagatg ggcatgaatt gatgagtgat gaactcaata ataggagggt180gcataactgc ctcctttgcc tccgttacca cctttgccac caccattctt accaccacct240cctttcccgg cattaggtcg tttgcagcat gggtattgct cgagatcgca ttcataaaag300agctaaaact ggcgctcgcc gggaacgaaa tcgcaagaag aggaaattcg agttgggtcg360cccggcagca ctgacaaagc ttgggcctaa acgtattcac accgttcgtg tcagaggtgg420aaatcttaag ttacgtgcaa tgcgcttgga tcagggaaac tttagctggc catctcaagc480tatttcacga aagacgaaaa ttattgatgt tgtgtataat gcgtcgaaca acgaacttgt540ccgaacgaag actcttgtca aaagcgcgat agttcaaatt gacggagctc cctttagaca600atggtttgag gctcactact tgaaagagct cggtcgccgc aaagttgtta gtaaaaaagg660acatacagta acacaagaaa atccagaaga agatattttg ttgaagaaaa gaagcaaatc720agcattgaaa aagtatgaaa ctcgtcaggc tttgcctcaa gcaaacgtag aagagtcttt780gaaggaagaa tttgttacag gtcgactctt agcttgtatt tcctctaaac ctggtcaggt840tggccgagca gacggctaca ttcttgaagg aaaagagttg gaattttact caaagaaatt900aaaagctaag aaagcaaaat aaatttctct tataataaac ttctgtagga c 951<210>10<211>624<212>DNA<213>日本血吸虫(Schistosoma japonicum)<400>10aattcggcac gagggtttgc gcgggttact tgttatcggg cgtgactgtt ggtagttttt 60caagccacta agttttaaga tgaagttcaa caaacatgtg agtagctctc gtcgcaaagc120tcgaaaacgt tacttcaacg ctccttctca tattcgccgg cgactaatga gcgctccttt180atcgaaggag ctgcgatcaa aatacaaagt cagaagtatg cccattcgca aaggggatga240agtacagatt gtgcgaggtg agcggaaaga ttgcaatcct gcaaaggtca ttcgtgtata300tcgcaaaaaa tatgtaattc acatcgagag gcttcagtgt cgtaagatga acggggcata360tgcacccatt ggcattcatc cgtccaatgt tgtcatacat aaattaaagt tggataagga420cagacgctca ttgttggagc gcaaggctgc cggtaaactc gctggacagg acaaaaacaa480atacacagaa gaaactattg agtcgtaatt tttattacat gcttttgtgg taaaaaaaaa540aaattaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa600aaaaaaaaaa aaaaaaaaaa aaaa 624<210>11<211>456<212>DNA<213>日本血吸虫(Schistosoma japonicum)<400>11tcggattgtg ttctagaaga catgtctgct cgtgcattta gaaagtttgc tcctctgttt60gaccgagtat tggtccaacg gtttgaagct gaaaccaaaa gtaagggtgg catcatgttg 120
ccagagaaag ctaagggcaa ggttcttgaa gcaactgtag ttgcacacgg tccaggagtg180aagaatgaga aaggtgaggt agttccagta tgcgtcactg ttggtgacaa ggtttttctc240cctgaatatg gtggaactaa agtcgtactt gaagataccg agtactttct tttccgagaa300tctgacatct tggctaaatt tgagaagtga ccggtgtctt agatctagtc tatctaccct360tgtacaatga ttattccaat tacatagttt ttcatctcta aaaaaaaaaa aaaaaaaaaa420aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaa 456<210>12<211>968<212>DNA<213>日本血吸虫(Schistosoma japonicum)<400>12agtgtgccat aaagcttgtc taactgccga aatgctgcat ctgaccaaat agctaaacgt 60cccatatgac cacccggagc cagcttaagg agatttaaag ctctcacgtt aatcaaaggt120gacacctgga atatttctaa aagcacgcga ttatcccatc atccttctta taaattataa180ttgggccacg tttttgaatt cgccttcgat ttctcatttc tccttttaat ttacaagtgt240gatcacgagg atgtcgttgt gccgacaaaa ctaccatgag gaatgtgaag ctggtattaa300caagcagatt aatatggagc tctatgctag ttatgtttat atgaccatgg cttttcattt360tcatcgtgat gacgtcgcgt tgaacgggtt ttataaattc tttcttaatg agtccgaaga420agagcgacaa catgcgataa agttaatgac gtaccaaaat atgcgaggtg ggcggattgt480gttgcaagat atatcagcac ctcctcaact ttcatggact tcaggtctcc atgccatgca540ggatgctttg gaattggaga aaaaagtcaa tcaaagtttg atggatttag ttgccgttgg600tgagaggcat cgggatacac acttttgtga ttttattgga aatgaatatt tggaaactca660aatacaatca atgaagaagc ttgctgatta cattacgaat ctcaaccgtg tgggcactgg720tttaggagaa tatactttcg ataaagaaac cttacacgga gaaagtgatt aatttcgtgc780tttcaagtag ctgtttgcgg attgtaataa actatttttt gtaaaataat ttatgtagat840tgatttacgg tattcttcag cgaaattcat caggttgcaa actagttaca tgtaatgtca900gaaaagtgtt tcattttttg ttattgatta ttacaatatg accaaactaa gtcttctttc960taaaaaaa 968<210>13<211>616<212>DNA<213>日本血吸虫(Schistosoma japonicum)<400>13atacgatgtg taccagagcc agttgttgga ggaccgtcat gcacaatgtt ggctatcaag 60tcgtaagtag tgtaccgatg tttagctcga aattctgaag acaaaagatc acctaaatca120atacttttaa tggggaaatt tacaattgtt ggatttttct ccagtgtgaa gatgttttta180acaaatctct tcatatagag aattaaataa ggaggaagtc gacgaagctc aaaccgacgc240aacgtagaat cacggtgagt tttatattcc ttctccgcaa ttccattaaa ttttgaaagt300agtgtagctg ttaatatcca aggtcgtaac atggcaaaga gcaagaacca tacaagtcat360aatcaaaata aaaagcaaca ccgcaacgga atcattagac cgaagaagca gcgatatccg420
tcacttaaag gcgttgaccc caagttcctt agaaatatga ggtttgccaa gaagcacaat 480aaacgtggga ataagaaggt acagcctgca gttactccaa cagtagcggg atgtaaataa 540tcgtggattg cactcgtaat atgtaaggaa taaatatact tatataaatc acaatgatcc 600agttcaaaaa aaaaac 616<210>14<211>432<212>DNA<213>日本血吸虫(Schistosoma japonicum)<400>14tttagcgcca ttgtttactt cttgtttact tgacacgtat tcacatggcc gataaatcac60ctgagaaaaa ccatatttca catcctccag cagtgaaagc tggagggatg cgaattgtga 120aacatgctaa agaacacgag gaaagtgtgt tatcgaagtc agatctagcc aaacaagcgg 180ttgaatataa ttctagtatt acaggtattt catcacacga tcttctcact aaggaagaac 240ttgaatctca tgatagatgt atcaaacttc atcaggaaaa gccgcttcca acagttgcaa 300aacctcataa caacgatagt acgagacgtg ttattcaaca acctttaaag tgaataacat 360gttgttttta aagaatgaag ttactctttt gtaaacttta ttttttaaaa taaaataagt 420taatatttat tt 432<210>15<211>1394<212>DNA<213>日本血吸虫(Schistosoma japonicum)<400>15attgaaacca ggtgtttcgc tcattacaag agtcgacgta cttgtccagg atttacgctg60tctgtttcgt tgcagcattg aacttttcct cctgatcgtt ttgctttatt ttttgcatag 120tgtattttta attgttgatc ggatacccaa aagcattgag cgtacttccc tcttagattg 180ctaagtttgc taggatcact cctgaaagtg atcagttggt tttcttcatt caaccttttc 240ccagagctta cgtgcctacc tcactactcc gagactttat tttctcatgt tttacagtat 300caagctgtga cacatgagac acagccacct ttttgtctgg tcgtgttacc ctctgagtac 360gttgtttgtt agtaaaagaa tgagtgtata aaacttgaaa aacgtattta agcagcatct 420taggtttcac acgtttgtaa ttctgtttgg ctttattcgg cttttgttta ttgttgtctt 480gttccgtttt ctattccatt tttctaacct ttagatcact tgactgaata cctttcaaga 540gcatcagaca cttggtccta catttgctgt tatcgctgag tacatttcta agtgcatctc 600gttttgagtc ttattctttc cctgatttct tcgtaaacac tgctctgcat tttatcccag 660tattggacac cttcatacta gtcgcacagt tgctaaaggc ttaacttcac atgtagcttt 720ccgtgatgac gtctgaaaca gtgaatataa tggacttaac cattccacaa ctccaagagc 780ttgcgaggca aatggagcaa aaaatacaat ttatatctgc ttccatacag caactaaagt 840cacttcaaac ccaattcatt tcttcgaaga attatctgtg cgaactgact ccagacaaag 900agaatagcga tattcttgtt cctttaactt caactttgtg tgtacctggg acactttccg 960acgcttcaca tatatttgtg gatatcggta cagggtatta tgttgaaatg accgtgctag 1020aagctgaaag tcactttgac cgtcgagtgg aatatatcaa taagcaaatc agaaagattt 1080
ttccagttct ggaggagaag actcgtgttc ataagtccat ttcagacact ttagatgcca 1140aaattcagga tttcattaag atccctccat caccgtgctc atagtatagt aagatgaact 1200tttaggactt gcacaaattt ccactattac tccgctgttt taccaatgaa gattgtgtga 1260aactaatgat gtagttggga tcatgccctg attattataa agtatgcaaa atttattagt 1320aactgagtct taaatctaaa tgtttatttg aataaatggt gaatcggatt ggtcaaaaaa 1380aaaaaaaaaa aaag1394<210>16<211>561<212>DNA<213>日本血吸虫(Schistosoma japonicum)<400>16tttggagaac tatttgcact tcatggtatt gtagaaacca atcgccacaa attcaagata60cggtttaagc gagattttgg caatgacgag ctcattttat ttgcgctact acgtgggaca 120caaagggaaa ttcgggcatg aattccttga gtttgagttc aggcctgaag gcaagttgcg 180ttatgctaac aactctaatt ataaaaatga cacaatgata cgcaaagagg catatgtgag 240cccttctgta atggaagaac tgaagagaat cgtgctggaa agcgatatta tgtcagaaga 300tgatgccaca tggcctgtac cggatagagt tggacgtcaa gagctcgaaa ttgtttgtgg 360tgatgaacac atatctttca caacctcgaa gataggctct ttgattgata ttaccaacag 420caaggatccg gatggactaa gaacgtatta ttacctggtg caagatctga agtgcctagt 480gttttcactg attggattgc attttaaaat aaagccaata taattcattc ccttcaaaaa 540aaaaaaaaaa aaaaaaaaaa a 561<210>17<211>1600<212>DNA<213>日本血吸虫(Schistosoma japonicum)<400>17atatttatgt tgtctaacgg tgttacttcg ggtaaaatgt catgtgcaga cagttgtcct60atggccgaga aagtacgctc tataatatat tggcggcatc caattataac aggcgttata 120ctgactcttt tactgattgt tgaatttagt tttatgtgtc tttctgcaat ctctttcatt 180gcttatatcg ggttatcact ccttgtttcc gtcaacttag tgcgtatgta ctatcatttt 240gtcgccaaaa cagaaaataa ttttgtcaga gaatacctgg aacgcgagat cactgtgcct 300caagacaaag cagaaaaagt ctctcgacgg ctgaccgaag ttattaacaa gggtttagta 360cgaactagag aaatattctt attaactgac cccgctgcat ctataaagtt tgctgtacta 420ctgtatctac tgacatacat tggagctcag ttcaactttc taactttatg catactgttc 480acttttggta tcttctgtgt gccaaagttt tatgaacgct atcaaccgga aattgataaa 540atactggagc tagcaaaatc taaaatagat atggtagtat cacaagtaaa atctcaaatt 600gaaaagctcc cgattattgg cggaggtggt cgtaaagaaa aagcacagta aattaaacgt 660ttccagctta tctaaatata tttcgtacca aattcgatga gcaacgatac aatttgttca 720aagcaattta caataaaatt cccttttctt cctataagta taataaatgc ataaaactgt 780aaagcatata aggaacaatt agttttacat tctttctctt ctccttgttt tgtgtgtgtt 840
tcctaccgat ttttatatat cgtgctttct gtctacaaaa attctctcta acgttgttat 900tcattcaaca aaaatttatg ataaattctt aattgtttct gtttgttgga gatttgggtt 960ttcattatag taatctgtgt attaatgaat agagacgatt attaaatatc tggtatcttt 1020caaaataaac aatgttcatc atccccctct gtcatatttt actgtgttta gattatccat 1080ttgttgtaac aacgctaatt tctttttctt tattcctaaa ggaattgtcc tatcatgtta 1140ctacaaaaca taattgaata ctgtttccat ttgataaaat tctatgtacg tattatctag 1200atgttactaa ctgcacaatg tttagcctac tttctttttt tgttctctcc gtggtaaatc 1260ctacttcttg ttgtccatta cagttttgct gcagtggttt ttttttcttg gttccattac 1320tgttatcatt tactattcaa atattgtgtt tggtattaca atgagcattt aatgacatta 1380acattattgt attggactaa ctaattggtc aactggttgt tttactacca ttattatata 1440gagaatgatg ttaatctctt ttgaacgtga tttatatttg catagttatt gttttcgctt 1500cggaattcca tagatatggt tcataaagct aatgagttac tttgaaacga agtaatctat 1560cttcgttaaa acaaaaactt acacttagta ttttccatct1600<210>18<211>928<212>DNA<213>日本血吸虫(Schistosoma japonicum)<400>18aggctaatca tctaagcatg cctaataagc aaaaaaaggt tattaaagcc aagaaaatta60aggaaatagt tcatcatgaa cctgatatac cagttcttcg aaaaggaaag gcttcaaaga 120ttgctcccct tccagatatt gccaagaaaa cgtctgttaa gaagaaagtt attaagaacc 180cactaatcca acgccgacct aaaaactttg gaatcggcca aaacattcag ccgaaacgga 240atctttaccg ctttgttaag tggcctaagt acatcgagct tcaaagaaaa aaggctatac 300ttaaaaagcg attgaagatc ccacctccaa tccatcaatt ttcccagaca ttggacagaa 360actcagctaa tcagatattt aacttagtta cgaagtatca accactgagt aaacaagcca 420agaaaatgct tttagttcat cgagcgcaac ttcgtgccga aggcaaaccc gatcaaccaa 480caccacgtaa acctactgtc cgtgctggta ttcgtgaagt aaccactgcc ataactcaga 540aaaaggcacg cctagttgtt atagcacacg acgtagagcc tattgaaata gtattatttc 600ttcctgctct gtgtagaaaa atgggcgttc cctactgtat agttaaaagt aaagcaaagc 660ttggcctcct tcttcacaga aagacgtgta cctgcctagc tttcacaaac attagagaag 720aggataaatc acgattgtcg aagatagttg aaatgggtga aaaacaatta caatgatcgc 780tttgaagaga ttcgcaaaca ttggggtggc ggaatcatgg gttcaaaatc tctcgctaaa 840caagctaaaa tcgagaaggc aaagagaaag gagcttcaaa ttaagcttgg ataatttatt 900aactttaata cagtttaatg gccaattt 928<210>19<211>1008<212>DNA<213>日本血吸虫(Schistosoma japonicum)<400>19cgttgcgcaa cgcatgggtc gcaggaaagg acctaagcat gatcatcagc tgaagagtcc60
aaacgtagaa cattccacgg aagaactaat tgcagaatcc tctgtcagtg aaaatttgac120tgaggcccct acgactaagt cacctgactt ggaggttact gcttgcagca cagagactat180tgacgtccta ggaaatggtc tcgtcgttat aaagacgtta agaaaaggtc tcggtaggga240aactcgacca agccacggtg atactgttgt tataaattac aaaggttggt tggaagatgg300gactcttgtc gatgatgttg aaaatgctaa aatagtactt ggcgatggtg atgttataca360tgcatttgat ctttctatcc ctttggccga acacaaagag acattcgaac tgattactga420tgcgcgattc gcctatggat ctcgtgggcg ggatccggat attccatcag gagcaaaatt480aacctatcac atagaaattc taaaagttga tgatcctccc tgttatgcca atatgccgaa540ttctgaaaga cttgcaattg ctaatcagaa aaaagataga ggcaactatt actacaggag600agaagaattt gcattcgcca tcgatagtta tagcaaagca ttgaaaattt tacaattacc660tacatcttct actcaatcat cgggggaaaa atcggatgta gattgtccga cagagcttat720taatgatgcg aagttaaaat tggaaaataa tcttgcagca gctcagttga aggttgaggc780gtttgatgcc gctataatgt cctgtgatac tgttttgcaa agcgaccccc aaaatattaa840ggctctgttt aggaagggaa aggctttatt agaaatgaat gaagtagatg atgcgatccc900aattcttcaa aaagtgctaa cattagctcc aggttcacag atggctagcg ttgagctagc960tcgtgctcga gctatacgac agaaggaacg cgaacattgg tcacgatc1008<210>20<211>821<212>DNA<213>日本血吸虫(Schistosoma japonicum)<400>20gttttttcaa tagtctgtga catgcgtgca aatggcgagc tgaaattatt ttctgtcgta 60ggccggaagc taccgacaac aaaggagcca aaacccgtcc cttacagaat gcggatattt120gctcctaact cggttgttgc aaaatcacgt ttctggtact tcattagaaa gctaaagaaa180atgaagaagg gtagcggtga gataatttct tgtgaaagaa tacacccacg aaagccattg240atggttaaaa atttcggtat ttggctaaga tatgatagtc gttcagggac acataacatg300tacaaggaat atcgtggttt gacggaggaa agtgcagtga ctcaattgta ccgtgagatg360ggtgctcgtc atcgtgctcg agcggagtct atccaggtaa ttaaagtgga ggcaatacca420gcatcgaagt gtcgtcgccc atatattact cagttccacg acagtaaact gaagtttcct480ctacctcatc gcgtggcccg tcctcttcat cgcccaaggt tcacaaccag aagacctaca540acagccttct agttttacaa taaatttgaa tggttacttt catttgttct tgattatgta600ttccgtgtta gattttgtat cctcactatt tcagtaaacg cgtattaatg cgaaattatg660gtttgttaat ggtgtataat ataggcagct cttcctcctc aactgtttta tctgccagct720tacgcgtatt caacatttaa ttggtacttt aatcatttca actgctttat ccagttgttt780gttcttatta ctttattatt ctgctaaagt atagtaaagg g821<210>21<211>943<212>DNA<213>日本血吸虫(Schistosoma japonicum)<400>21
gtgcaatctt tcgaatatgc ctacattaag ctgtggatat aagtgtttgc aattcttgtt 60aatcaccttt aatatccttg tttttgcatg tgggattgct ttaatagtta ttgggagtct 120ctctcaagtg gctgttaata actactcgac gggaatcgat tcaaatatca aaggactcgt 180aatatttatc ataatactcg gatgctttct cttcttgctc ggatttcttg gattttgcgg 240ggcatgcaca aaaaatactt gctgtttaac aatatatgct attcttttgt ccgttatggt 300tgcggctgaa ataggggtag gaatagccgc tgccgtccta aaagaagatg tgaaaacaag 360ctttgtgtct gtggttagga actctgtttc ggaatatagt aaaaatcctg atattcagaa 420tctttttgac aagatacaaa cagagtttaa atgttgtggt tccgagtcat ctatcgatta 480tacatcaact ggacaatccg ttccatcctc ctgtacggat tcagatactg ggcttgcgta 540ccaagaaggg tgctcaaaca aaataattgc cttcttcgaa aagtatctta ttgctgttgt 600ggtggcagca ttcgcattcg ctatccttca aatattaagt attgttttta ctgtctgtat 660tattcgagct attaaatctg gtgatactga gtgaaactaa aaatcataaa cgtgtaaaaa 720ctagctttga tttttcataa acttccaaat taatgaaatt aaactttttt tcattgtatc 780tttattttgc tgtcatttaa cacttggaaa acggagccat attttttatt aaaacatata 840ctagaaaact acttatgaat attttatggt tactggacta acggttataa taatgacaaa 900attaaataaa atttgaatac tttcttaaaa aaaaaaaaaa aaa943<210>22<211>812<212>DNA<213>日本血吸虫(Schistosoma japonicum)<400>22taaaaaagaa acaatgaaga ttttatttat ttgtctgcta cttattacac tacttactat 60gactattcaa tataatctaa atcgaaaata cactaatcat tatcggagac aaccaaataa 120tccttataat cgtaataata atggtaataa taattatgat gataacaaca ataatgacaa 180caaaaatatg tatgataagt atagtgataa tagtaaatac ggtaattatt acccaactaa 240taatgattat aataaacaga aaccaaaata tccacgtctt accagaagat acagtatgga 300aggtgaagaa aagcatgcgc atcaaaaaca tggtgaacgt catttaaaat ctattggtga 360attatttggt tttactaata caaatcaagg ttcagactac gcattaacaa caacatttca 420taaaggtagt atcgctgcta aaaatggtaa acaaaagcat ttttctactt atgaaacaga 480aggtcaagtg aaacactatc agaatacaca tggtcaggct gaattcaatc taaatgctaa 540actatatggt caagaaaagt ataatgatca gaaggaaatg aaagcaacag taaaaatgga 600agattatgat aatacacctg acgacgaagc aaattactca ccagtcggac atccatatgg 660taaattagaa ggctactaat tatgattaaa aaaaacaagc gctcataata aatatgaaat 720tataagtcag ttgaattgaa tgtatataac ttccatatta ttctcttttg agtcatgaca 780tgttgaaaat cgaataaatc ttataataag at812<210>23<211>1535<212>DNA<213>日本血吸虫(Schistosoma japonicum)<400>23
aggagctgca gttctcacat ctactgcaag tctaggctgt gatccatttt tactgccccc 60ggatcatcga agacttacaa accatcaacg atcatttgcc gctggctatt caagtgatca120cttggctggt ttgaatgtct ttcaagtatt ggcttctgaa cgtaaccgtc ggggtgatga180gtcagctgat tcattttgtg accaacatgg gttcaatagc tctgcgttac gaattattga240tgatgctgca aaccaactta gagctatcct tatcaatttg ggttttcctg aagaatcatt300gtctgatgca ccggttgatt tcaacgttaa acacaatatt gattgcgata taattaatac360tttgctggtg tctggattgt atccaaatat ctgttttcat tcagataaac gcaaattact420cacaactgaa ggtactctgg ctctcataca caaaggttct gtcaattgca tcaaagatat480caagttcact tacccatttt ttgtttttga tgaaaaaatt cgtactcagg ccgtttcatg540caaaggtctc acaatggtaa atccaattca actgatgctc tttggttgtc gctcagctgt600ctggaggaat agtgattcta atgataaagg aatagttgtt attgatgatt ggattccgtt660ccgcatgagc tttacttcag ccgctcggat ttttgctttt cgaccagcat tggaagcatt720gcttgtccga acttgcttgc gaccagaagg tattgctgat cttagagagg aagataagcg780tgttattaat gtattgcgtc aactatgctc accagaagct ttaaatccaa acgctagttt840cgatgaagca agcagttacg actctccacc gagaaaaaag tttgctacag ctggtaatta900tccgttactt tctggtcgat cgcatcgcac tgtgggttat gagaattcat gctcagagaa960acaaggtttt ggacgttatg ctaattgtcc tagtcggttt ggtgtctcag aacaagcatt 1020cgatggtcag tgtggttttt ggcaggaccc tctttctcaa ttttcaaatc agaggtatga 1080ccaatcatct gtcaacagat ttcctgggaa ctacacaagt atacggtcgt ttggcgctag 1140cggtatggga aataatctta atggatcatc ttgttactcg tatggagtta ctccttttgg 1200tggatacccg aatcaacagc cacagtttac aaatgatagg tttattcaac aaagtcaaag 1260gccagtaaat agacctcatt ggggcatacg ttattagctt taatttttga aactgttttt 1320aattgtatcc aagtattaga ttcttctcct aaactctgag tatttattta tttggactca 1380tttctatatg tactgtattg atatccccat ctttctaaca tgctaaactt tgtttcttgt 1440tgactaactt taatgacctg gtgtctaaac cttcatttct acgcactgtt ttgttattgc 1500catctaagtt acaataataa aatcgttcta agttt 1535<210>24<211>1055<212>DNA<213>日本血吸虫(Schistosoma japonicum)<400>24acctgatatg ccgaattatg aagtattatc tccacaggtt gtttgcccaa catgcaccgg 60agcaggctat gtgagaagag ggctgagtat gtggctttaa tccctttgac agaccatcgt120ctccggccgc gtagaatctg tcttaaattg tttggctctg tagtgatttg ctccttagtt180tttactctct tactgatttt tctctttcca cgaacaatat atttgcgaag ttcagtttac240aagttaatta ctgtcagtgc catcgccaat atgcaagata atgtgattga tctcactgtt300attaaccaga ttaatgtttc aaatccaaac tttgttcctg ccatggtaaa taaggtcatc360atctctccga aataccgttc agtggatcta caagagactg taaacaattc catccatatg420ttaccacttc gttccgaaac tttactgaat gtcagcacca agttacgctt cgagcataag480cattttgatg tcattaaatt gtgcttcatg aaattgcctt acttccacga tatatacgta540acttttacat ttcatatcac ttacatgttg ttatggcaaa gttttgatgc atatcttaca600attatgccat tggttgattg ttatccaact aatataaatt catccatgtt gatggtgtct660
agttaaatgg tttatcattc ttgctaatca tcattgattc ttttttatat tgaatatgat720ttaaagagat ttttataatt tagactttct atctacgcaa ttattttcta ttgaaagtca780ctatgcttat tgacttttat ttgaatattt tatttttctc attgtgattt ttaatctatc840attgttcagt tgattagaaa gtgatagcga gcgcgagaga gagaacgttt tttccacatc900taattcctat ttgttttgca tttgttatta attgtatgaa tacaaaatat gaagtcatta960tgcttttgaa ttgttattat tgtattccaa ctgtttctac tatttcaatg tagtggtcgg 1020atttatctat gttaataaac caattgagtt atact 1055<210>25<211>448<212>PRT<213>日本血吸虫(Schistosoma japonicum)<400>25MRECISLHVG QAGAQMGDSC WELYCLEHGI QPDGKVPSDS TIKNDNDSFS TFFSETSYGR 60YVPRAVFVDL EPSVIDRLRT GAYKQLFHPE QMISGKEDAA NNYARGHYTV GKELVDEVLD120RIRKLSERCT GLQGFLVFHS FGGGTGSGFT SLLMERLSVD FGKKAKLEFA VYPAPHISTA180VVEPYNSILT THTTLEHSDC AFMVDNEAIY EICRRNLTID RPTYCNLNRL IGQIVSSITA240SLRFSGALNV DLNEFQTNLV PYPRIHFPLT TYAPIISAEK AFHEQLSVSE ITNSCFEPAN300QMVKCDPRNG KYMACCLLYR GDVVPKDVNA AIANIKTRRS IQFVDWCPTG FKVGINYQPP360TVVPGGDLAK VQRAVCMLSN TTAISEAWAR LNRKFDLMYA KRAFVHWYVG EGMEEGEFSE420AREDLAALEK DYEEVASDTV DAQSDDGF 448<210>26<211>387<212>PRT<213>日本血吸虫(Schistosoma japonicum)<400>26MMPAYTNGGN EELETFLFTS ESVGEGHPDK LCDQISDAIL DEFLKQDPDA KVACESFVKS 60GLIIVGGEIS SNAVIDIQKT VRNVVKDIGY DDTRKGLDYK NCNILAVIEQ QSADIAKGVH120EGRNVEDIGA GDQGLMFGYA TDETEECMPL TVTLSHALNK RLAMCRRNGI LPWAGPDSKT180QVTVEYLNDH GACRPLRVHT VVISAQHDEH VTLEKVRADL MEYVIKEEIP ANLLDEKTKY240HLNPAGRFVI GGPQSDAGLT GRKIIVDTYG GWGAHGGGAF SGKDYSKVDR SAAYAARWVS300GLCRRVLVQL SYAIGIAEPL SINVNSYGTG KISDKKLLDV IMKNFDLRPG IIVKDLDLRR360PRYLQTSVYG HFGRPEFPWE QCRKLKL387<210>27<211>294<212>PRT<213>日本血吸虫(Schistosoma japonicum)<400>27MNHTRALHFV FKVANRNETI DFYKRILGMK VLRHEEFSEG CKASCNGPYD GKWSKTMIGY 60
GPEDNHFVIE LTYNYGVGAY QLGNDFQYIR IHNKEAYSRI INGSWPITSE ESDSVKVKAP120GGYTFWVHNS DSDGDPVRMV ALSSSDIKRS IDYWSHKLHM TLVSSSSNEA VFYYSPNQCN180LKLISIQKPV DHASAFGRIA FACPSAQLPI LQELMDKENE TILTRLVSLD TPGKATVQVI240ILADPDGHEI CFVGDEAFRQ LSQVDSQADS LLQTAITSDK SDEWYNKHGG KTTK 294<210>28<211>65<212>PRT<213>日本血吸虫(Schistosoma japonicum)<400>28MPTKVLRVTT RKTPCGEGSK TWDRFQLRIH KRVIDLLSTD EATLQLKHIY LEPGVNCDVK 60VSGDL 65<210>29<211>248<212>PRT<213>日本血吸虫(Schistosoma japonicum)<400>29MNLYCIIRIG DARFETQTSS RSGKHPIWNE TFRCWLHDGV DKLSLEIVSE NIFSSSQVAH 60AVIAFPRSLY DGIALNQWFE LSGQQGENLE GSINLIFQCR KVPVNLPLLQ STPLYCQPRL120GVVGDMQVLP KPISTEEVQS IHEIFPSVEE DTIRSLLATH HGNREIVISE LLLWHRKLEI180VCSDCDVFKS FSNNVIFLFI CYWNDFSSEQ RFPLTALTCY YPVLLQPPLS NFLTKIKQRY240DHFYTDFI 248<210>30<211>431<212>PRT<213>日本血吸虫(Schistosoma japonicum)<400>30MNKDAVDMSE TPKQSEEEYL KDFIDLSPSG DRGILKKVVR EGYSDIKPCD GDTVIVHYVG 60TNFGGEKHGE VFDSSRARNE KFEFTIGKGS VIKAWDIGVA TMRLGEVCEL IASPEYAYMD120GKSLKFEVEL FETMGSDVSR NKDGSIRKSI IKKGRDIHNP VAGAEATIVF RNLLNLTEET180EVTYCVGDPP SNVPDELDQS VRHMNTGEVS RIVVYKDGHS LTSGDSDKDR IVYELTLKSF240EKTKHLSGIT SFPERIAYAN TLKEKANNFL KDSKFDSAIE LYKRLDDELQ YVVANGPTEQ300KELSGVTVAV QLNLALVYLK LCKPDKCIEF CKKVLDNFSD NEKALFRIGQ AHLLRKDHEE360AVVYFKRIVT KNPNNASAVK QVQICEEEIR RAKDIERKKF RHIFERCKDT GLNDAQNHED420GVVVNDEKSAL 431<210>31<211>552<212>PRT
<213>日本血吸虫(Schistosoma japonicum)<400>31RFHLVMLTSG DSDLVGEAVK FHKPGENYNS PGYVVTKNTL RLLKEHLKQT GGKVVTRFPP 60EPNGILHIGH AKAINFNFGY AMKNGGITYL RYDDTNPEKE EAEFFEAIED MVRWLGFTPY120KITYASDNFH QLYEWSVEEI RGFNPPPSPW RERPVEESLR LFEDMKNGKI DEGMATLRLK180VTLGDGKVDP VAYRIKMTPH HRTGNEWCIY PTYDYTHCLC DSIENITHSL CTKEFQSRRP240AYYWLCNALD VYCPVQWEYG RLNLYYSVVS KRKILKLIEA GIVSDWDDPR LVTLTALRRR300GFPPQAINMF CERIGVTMAQ TILDPSALDA CVRDYLNDHA PRVMAVLEPV LVTITNWFEL360YGDKSSIELT VADFPAVPDS KTHSITLQHD LYIEKTDFQE VAEKGYRRLT PTQAVGLRHT420GLVLEFSDFK KDSNGEIVHL FVKAKKVEDS IKPKAFIHWV SNPLQFEARL YDRLFTVKEP480ESAKEGFLSV INKDSLVVLS DALIEQSVMN AKVYTAYQFE RIGFFSVDPD TTSKRMVFNR540TVALKVDPGK TI552<210>32<211>169<212>PRT<213>日本血吸虫(Schistosoma japonicum)<400>32MRVFKDAISG DEMFSDSHSP QLINDVVYEV DANFITVSNG LDSKLIAANP SGEEGQEEVS60DSTERVIDLV HASRLVSTSF DKKSYRAYLK GYLKAIKERL QKENPERVSI FESRINEYMV 120NVFKNFDDYE HYIGESMNPD GMVALMNFRE NGVTPYFVFL KDGLIEEKY 169<210>33<211>217<212>PRT<213>日本血吸虫(Schistosoma japonicum)<400>33MGIARDRIHK RAKTGARRER NRKKRKFELG RPAALTKLGP KRIHTVRVRG GNLKLRAMRL60DQGNFSWPSQ AISRKTKIID VVYNASNNEL VRTKTLVKSA IVQIDGAPFR QWFEAHYLKE 120LGRRKVVSKK GHTVTQENPE EDILLKKRSK SALKKYETRQ ALPQANVEES LKEEFVTGRL 180LACISSKPGQ VGRADGYILE GKELEFYSKK LKAKKAK217<210>34<211>142<212>PRT<213>日本血吸虫(Schistosoma japonicum)<400>34MKFNKHVSSS RRKARKRYFN APSHIRRRLM SAPLSKELRS KYKVRSMPIR KGDEVQIVRG60ERKDCNPAKV IRVYRKKYVI HIERLQCRKM NGAYAPIGIH PSNVVIHKLK LDKDRRSLLE 120RKAAGKLAGQ DKNKYTEETI ES142
<210>35<211>109<212>PRT<213>日本血吸虫(Schistosoma japonicum)<400>35SDCVLEDMSA RAFRKFAPLF DRVLVQRFEA ETKSKGGIML PEKAKGKVLE ATVVAHGPGV 60KNEKGEVVPV CVTVGDKVFL PEYGGTKVVL EDTEYFLFRE SDILAKFEK109<210>36<211>173<212>PRT<213>日本血吸虫(Schistosoma japonicum)<400>36MSLCRQNYHE ECEAGINKQI NMELYASYVY MTMAFHFHRD DVALNGFYKF FLNESEEERQ 60HAIKLMTYQN MRGGRIVLQD ISAPPQLSWT SGLHAMQDAL ELEKKVNQSL MDLVAVGERH120RDTHFCDFIG NEYLETQIQS MKKLADYITN LNRVGTGLGE YTFDKETLHG ESD 173<210>37<211>69<212>PRT<213>日本血吸虫(Schistosoma japonicum)<400>37MAKSKNHTSH NQNKKQHRNG IIRPKKQRYP SLKGVDPKFL RNMRFAKKHN KRGNKKVQPA 60VTPTVAGCK 69<210>38<211>102<212>PRT<213>日本血吸虫(Schistosoma japonicum)<400>38MADKSPEKNH ISHPPAVKAG GMRIVKHAKE HEESVLSKSD LAKQAVEYNS SITGISSHDL 60LTKEELESHD RCIKLHQEKP LPTVAKPHNN DSTRRVIQQP LK 102<210>39<211>152<212>PRT<213>日本血吸虫(Schistosoma japonicum)<400>39
MTSETVNIMD LTIPQLQELA RQMEQKIQFI SASIQQLKSL QTQFISSKNY LCELTPDKEN 60SDILVPLTST LCVPGTLSDA SHIFVDIGTG YYVEMTVLEA ESHFDRRVEY INKQIRKIFP120VLEEKTRVHK SISDTLDAKI QDFIKIPPSP CS 152<210>40<211>146<212>PRT<213>日本血吸虫(Schistosoma japonicum)<400>40MTSSFYLRYY VGHKGKFGHE FLEFEFRPEG KLRYANNSNY KNDTMIRKEA YVSPSVMEEL 60KRIVLESDIM SEDDATWPVP DRVGRQELEI VCGDEHISFT TSKIGSLIDI TNSKDPDGLR120TYYYLVQDLK CLVFSLIGLH FKIKPI 146<210>41<211>214<212>PRT<213>日本血吸虫(Schistosoma japonicum)<400>41MLSNGVTSGK MSCADSCPMA EKVRSIIYWR HPIITGVILT LLLIVEFSFM CLSAISFIAY 60IGLSLLVSVN LVRMYYHFVA KTENNFVREY LEREITVPQD KAEKVSRRLT EVINKGLVRT120REIFLLTDPA ASIKFAVLLY LLTYIGAQFN FLTLCILFTF GIFCVPKFYE RYQPEIDKIL180ELAKSKIDMV VSQVKSQIEK LPIIGGGGRK EKAQ214<210>42<211>252<212>PRT<213>日本血吸虫(Schistosoma japonicum)<400>42MPNKQKKVIK AKKIKEIVHH EPDIPVLRKG KASKIAPLPD IAKKTSVKKK VIKNPLIQRR 60PKNFGIGQNI QPKRNLYRFV KWPKYIELQR KKAILKKRLK IPPPIHQFSQ TLDRNSANQI120FNLVTKYQPL SKQAKKMLLV HRAQLRAEGK PDQPTPRKPT VRAGIREYTT AITQKKARLV180VIAHDVEPIE IVLFLPALCR KMGVPYCIVK SKAKLGLLLH RKTCTCLAFT NIREEDKSRL240SKIVEMGEKQ LQ252<210>43<211>331<212>PRT<213>日本血吸虫(Schistosoma japonicum)<400>43MGRRKGPKHD HQLKSPNVEH STEELIAESS VSENLTEAPT TKSPDLEVTA CSTETIDVLG 60
NGLVVIKTLR KGLGRETRPS HGDTVVINYK GWLEDGTLVD DVENAKIVLG DGDVIHAFDL120SIPLAEHKET FELITDARFA YGSRGRDPDI PSGAKLTYHI EILKVDDPPC YANMPNSERL180AIANQKKDRG NYYYRREEFA FAIDSYSKAL KILQLPTSST QSSGEKSDVD CPTELINDAK240LKLENNLAAA QLKVEAFDAA IMSCDTVLQS DPQNIKALFR KGKALLEMNE VDDAIPILQK300VLTLAPGSQM ASVELARARA IRQKEREHWS R 331<210>44<211>176<212>PRT<213>日本血吸虫(Schistosoma japonicum)<400>44MRANGELKLF SVVGRKLPTT KEPKPVPYRM RIFAPNSVVA KSRFWYFIRK LKKMKKGSGE 60IISCERIHPR KPLMVKNFGI WLRYDSRSGT HNMYKEYRGL TEESAVTQLY REMGARHRAR120AESIQVIKVE AIPASKCRRP YITQFHDSKL KFPLPHRVAR PLHRPRFTTR RPTTAF176<210>45<211>225<212>PRT<213>日本血吸虫(Schistosoma japonicum)<400>45MPTLSCGYKC LQFLLITFNI LVFACGIALI VIGSLSQVAV NNYSTGIDSN IKGLVIFIII 60LGCFLFLLGF LGFCGACTKN TCCLTIYAIL LSVMVAAEIG VGIAAAVLKE DVKTSFVSVV120RNSVSEYSKN PDIQNLFDKI QTEFKCCGSE SSIDYTSTGQ SVPSSCTDSD TGLAYQEGCS180NKIIAFFEKY LIAVVVAAFA FAILQILSIV FTVCIIRAIK SGDTE225<210>46<211>221<212>PRT<213>日本血吸虫(Schistosoma japonicum)<400>46MKILFICLLL ITLLTMTIQY NLNRKYTNHY RRQPNNPYNR NNNGNNNYDD NNNNDNKNMY 60DKYSDNSKYG NYYPTNNDYN KQKPKYPRLT RRYSMEGEEK HAHQKHGERH LKSIGELFGF120TNTNQGSDYA LTTTFHKGSI AAKNGKQKHF STYETEGQVK HYQNTHGQAE FNLNAKLYGQ180EKYNDQKEMK ATVKMEDYDN TPDDEANYSP VGHPYGKLEG Y221<210>47<211>247<212>PRT<213>日本血吸虫(Schistosoma japonicum)<400>47
MVNPIQLMLF GCRSAVWRNS DSNDKGIVVI DDWIPFRMSF TSAARIFAFR PALEALLVRT 60CLRPEGIADL REEDKRVINV LRQLCSPEAL NPNASFDEAS SYDSPPRKKF ATAGNYPLLS120GRSHRTVGYE NSCSEKQGFG RYANCPSRFG VSEQAFDGQC GFWQDPLSQF SNQRYDQSSV180NRFPGNYTSI RSFGASGMGN NLNGSSCYSY GVTPFGGYPN QQPQFTNDRF IQQSQRPVNR240PHWGIRY 247<210>48<211>204<212>PRT<213>日本血吸虫(Schistosoma japonicum)<400>48MHRSRLCEKR AEYVALIPLT DHRLRPRRIC LKLFGSVVIC SLVFTLLLIF LFPRTIYLRS60SVYKLITVSA IANMQDNVID LTVINQINVS NPNFVPAMVN KVIISPKYRS VDLQETVNNS 120IHMLPLRSET LLNVSTKLRF EHKHFDVIKL CFMKLPYFHD IYVTFTFHIT YMLLWQSFDA 180YLTIMPLVDC YPTNINSSML MVSS 20权利要求
1.一类日本血吸虫雌性成虫特异蛋白，其特征在于，选自下组(a)具有SEQ ID No.25～SEQ ID No.48所示氨基酸序列的蛋白；(b)由(a)的蛋白发生一个或几个氨基酸置换、缺失或插入而形成的具有(a)蛋白相同活性的蛋白。
2.如权利要求1所述的蛋白，其特征在于，它具有SEQ ID No.25～SEQ ID No.48所示的氨基酸序列。
3.一类日本血吸虫雌性成虫特异蛋白基因，其特征在于，它编码权利要求1所述的蛋白。
4.如权利要求3所述的基因，其特征在于，选自下组(a)SEQ ID No.1～SEQ ID No.24所示的核苷酸序列；(b)SEQ ID No.1～SEQ ID No.24所示的核苷酸序列中每条序列的互补序列；(c)与SEQ ID No.1～SEQ ID No.24所示的核苷酸序列中每条序列有至少70％同源性、且编码相同功能蛋白质的核苷酸序列。
5.如权利要求3或4所述的基因，其特征在于，它具有SEQ ID No.1～SEQ ID No.24所示的核苷酸序列。
6.一类日本血吸虫病防治疫苗，其特征在于，所述疫苗以权利要求1所述的蛋白为免疫原。
7.一类能与权利要求1所述的蛋白特异性结合的抗体。
8.权利要求1所述的蛋白在制备预防和治疗日本血吸虫病的药物中的应用。
全文摘要
本发明公开了日本血吸虫(Schistosoma japonicum)雌性成虫特异蛋白及其基因序列，还公开了与该蛋白特异结合的抗体，以及把雌性成虫特异蛋白作为免疫原制备所得的日本血吸虫病防治疫苗，此外，本发明还公开了日本血吸虫雌性成虫特异蛋白在制备预防和治疗日本血吸虫病的药物中的应用。运用本发明的蛋白，可抑制血吸虫雌性成虫的产卵，从而治疗血吸虫病、防止血吸虫的扩散。
文档编号A61K38/17GK1986567SQ20051011173
公开日2007年6月27日 申请日期2005年12月20日 优先权日2005年12月20日
发明者韩泽广, 刘锋, 王志勤, 胡薇, 冯正 申请人:上海人类基因组研究中心, 中国疾病预防控制中心寄生虫病预防控制所