專(zhuān)利名稱(chēng)：血吸蟲(chóng)雌蟲(chóng)特異蛋白基因、蛋白及其應(yīng)用的制作方法
技術(shù)領(lǐng)域：
本發(fā)明涉及生物基因領(lǐng)域，尤其涉及日本血吸蟲(chóng)雌性成蟲(chóng)特異蛋白質(zhì)的基因核酸序列、以及特異蛋白質(zhì)的氨基酸序列；此外，本發(fā)明還涉及日本血吸蟲(chóng)雌性成蟲(chóng)特異蛋白的應(yīng)用。
背景技術(shù)：
血吸蟲(chóng)病由血吸蟲(chóng)的感染引起，在世界上有76個(gè)國(guó)家，受感染者約1.5億。致病的血吸蟲(chóng)有3種，既日本血吸蟲(chóng)(Schistosoma japonicum)，曼氏血吸蟲(chóng)(S.mansoni)和埃及血吸蟲(chóng)(S.haematobium)。在我國(guó)流行的是日本血吸蟲(chóng)。血吸蟲(chóng)病的流行給人民身體健康和經(jīng)濟(jì)發(fā)展都造成巨大的損失，因此，防治血吸蟲(chóng)病、開(kāi)發(fā)研制治療血吸蟲(chóng)病和抗血吸蟲(chóng)的藥物具有巨大的社會(huì)和經(jīng)濟(jì)效益。
研制新的治療血吸蟲(chóng)病和抗血吸蟲(chóng)的藥物，首先必須找到合適的藥物作用靶點(diǎn)。血吸蟲(chóng)的蟲(chóng)卵是造成血吸蟲(chóng)病的最主要原因，肉芽腫就是由蟲(chóng)卵造成的。此外，蟲(chóng)卵排出體外進(jìn)入外界環(huán)境，污染環(huán)境后會(huì)制造更多的感染。所以，治療血吸蟲(chóng)病、防止血吸蟲(chóng)的擴(kuò)散，一個(gè)關(guān)鍵的措施是抑制血吸蟲(chóng)雌蟲(chóng)成蟲(chóng)的產(chǎn)卵。這可以通過(guò)干擾雌蟲(chóng)性成熟，干擾雌、雄蟲(chóng)的配對(duì)，或者抑制產(chǎn)卵相關(guān)的生理途徑來(lái)實(shí)現(xiàn)。而雌蟲(chóng)特異表達(dá)的蛋白質(zhì)，很可能與雌蟲(chóng)性成熟、雌雄蟲(chóng)的配對(duì)，或者產(chǎn)卵密切相關(guān)。通過(guò)大規(guī)模的基因表達(dá)譜學(xué)研究，發(fā)現(xiàn)了日本血吸蟲(chóng)雌性成蟲(chóng)的特異蛋白質(zhì)，這些蛋白質(zhì)，都是可能的藥物作用靶點(diǎn)和候選疫苗。
在本發(fā)明之前，還沒(méi)有出現(xiàn)涉及本發(fā)明的日本血吸蟲(chóng)雌性成蟲(chóng)特異蛋白的公開(kāi)報(bào)道。

發(fā)明內(nèi)容
本發(fā)明要解決的技術(shù)問(wèn)題之一是提供一類(lèi)日本血吸蟲(chóng)雌性成蟲(chóng)特異蛋白，及其基因的核酸序列。
本發(fā)明要解決的技術(shù)問(wèn)題之二是提供一類(lèi)日本血吸蟲(chóng)病防治疫苗，該疫苗以日本血吸蟲(chóng)雌性成蟲(chóng)特異蛋白為免疫原。
本發(fā)明要解決的技術(shù)問(wèn)題之三是提供一類(lèi)與日本血吸蟲(chóng)雌性成蟲(chóng)特異蛋白結(jié)合的抗體。
本發(fā)明要解決的技術(shù)問(wèn)題之四是提供日本血吸蟲(chóng)雌性成蟲(chóng)特異蛋白在制備預(yù)防和治療日本血吸蟲(chóng)病的藥物中的應(yīng)用。
為了解決上述技術(shù)問(wèn)題，本發(fā)明通過(guò)如下技術(shù)方案實(shí)現(xiàn)在本發(fā)明的一個(gè)方面，提供了一類(lèi)日本血吸蟲(chóng)雌性成蟲(chóng)特異蛋白，包括(a)具有SEQ ID No.24～SEQ ID No.46所示氨基酸序列的蛋白；(b)由(a)的蛋白發(fā)生一個(gè)或幾個(gè)氨基酸置換、缺失或插入而形成的具有(a)蛋白相同活性的蛋白。
較佳地，所述蛋白是具有SEQ ID No.24～SEQ ID No.46氨基酸序列的蛋白。
在本發(fā)明的另一方面，提供了一類(lèi)編碼日本血吸蟲(chóng)雌性成蟲(chóng)特異蛋白的基因，包括(a)SEQ ID No.1～SEQ ID No.23所示的核苷酸序列；(b)SEQ ID No.1～SEQ ID No.23所示的核苷酸序列中每條序列的互補(bǔ)序列；(c)與SEQ ID No.1～SEQ ID No.23所示的核苷酸序列中每條序列有至少70％同源性、且編碼相同功能蛋白質(zhì)的核苷酸序列。
較佳地，所述基因具有SEQ ID No.1～SEQ ID No.23所示的核苷酸序列。
在本發(fā)明的另一方面，還提供了一類(lèi)日本血吸蟲(chóng)病防治疫苗，該疫苗以日本血吸蟲(chóng)雌性成蟲(chóng)特異蛋白為免疫原。
在本發(fā)明的另一方面，還提供了一類(lèi)與日本血吸蟲(chóng)雌性成蟲(chóng)特異蛋白結(jié)合的抗體。
此外，本發(fā)明還提供了日本血吸蟲(chóng)雌性成蟲(chóng)特異蛋白在制備預(yù)防和治療日本血吸蟲(chóng)病的藥物中的應(yīng)用。
在本發(fā)明中，術(shù)語(yǔ)“日本血吸蟲(chóng)雌性成蟲(chóng)特異蛋白”指具有日本血吸蟲(chóng)雌性成蟲(chóng)特異蛋白活性的SEQ ID No.24～SEQ ID No.46所示氨基酸序列的蛋白質(zhì)。該術(shù)語(yǔ)還包括具有與日本血吸蟲(chóng)雌性成蟲(chóng)特異蛋白質(zhì)相同功能的、SEQ ID No.24～SEQ IDNo.46序列的變異形式。這些變異形式包括(但并不限于)若干個(gè)(通常為1-50個(gè)，較佳地1-30個(gè)，更佳地1-20個(gè)，最佳地1-10個(gè))氨基酸的缺失、插入和/或取代，以及在C末端和/或N末端添加一個(gè)或數(shù)個(gè)(通常為20個(gè)以?xún)?nèi)，較佳地為10個(gè)以?xún)?nèi)，更佳地為5個(gè)以?xún)?nèi))氨基酸。例如，在本領(lǐng)域中，用性能相近或相似的氨基酸進(jìn)行取代時(shí)，通常不會(huì)改變蛋白質(zhì)的功能。又比如，在C末端和/或N末端添加一個(gè)或數(shù)個(gè)氨基酸通常也不會(huì)改變蛋白質(zhì)的功能。該術(shù)語(yǔ)還包括日本血吸蟲(chóng)雌性成蟲(chóng)特異蛋白的活性片段和活性衍生物。
在本發(fā)明中，術(shù)語(yǔ)“編碼日本血吸蟲(chóng)雌性成蟲(chóng)特異蛋白的基因序列”指編碼具有日本血吸蟲(chóng)雌性成蟲(chóng)特異蛋白活性的核苷酸序列，如SEQ ID No.1～SEQ ID No.23所示的核苷酸序列及其簡(jiǎn)并序列。該簡(jiǎn)并序列是指，位于SEQ ID No.1～SEQ ID No.23序列中，有一個(gè)或多個(gè)密碼子被編碼相同氨基酸的簡(jiǎn)并密碼子所取代后而產(chǎn)生的序列。由于密碼子的簡(jiǎn)并性，所以與SEQ ID No.1～SEQ ID No.23所示核苷酸序列的同源性低至約70％的簡(jiǎn)并序列也能編碼出SEQ ID No.24～SEQ ID No.46所示的氨基酸序列。
在本發(fā)明中，可以使用一系列本領(lǐng)域已知的方法來(lái)制備針對(duì)日本血吸蟲(chóng)雌性成蟲(chóng)特異蛋白的抗體。例如，將提純的日本血吸蟲(chóng)雌性成蟲(chóng)特異蛋白產(chǎn)物或它的抗原片段注射入動(dòng)物體內(nèi)以產(chǎn)生多克隆抗體。同樣，表達(dá)日本血吸蟲(chóng)雌性成蟲(chóng)特異蛋白或它的抗原片段的細(xì)胞也可以用來(lái)對(duì)動(dòng)物致免疫而產(chǎn)生抗體。本發(fā)明制備的抗體也可以是單克隆抗體，這些單克隆抗體可用雜交瘤技術(shù)制備。
應(yīng)用本發(fā)明的日本血吸蟲(chóng)雌性成蟲(chóng)特異蛋白，可抑制血吸蟲(chóng)雌性成蟲(chóng)的產(chǎn)卵，從而治療血吸蟲(chóng)病、防止血吸蟲(chóng)的擴(kuò)散，該類(lèi)蛋白在血吸蟲(chóng)病防治和治療中有著重要的作用。


圖1是用RT-PCR實(shí)驗(yàn)驗(yàn)證日本血吸蟲(chóng)成蟲(chóng)特異表達(dá)基因的圖；圖2是圖1的數(shù)據(jù)分析圖。
具體實(shí)施例方式
下面結(jié)合附圖和實(shí)施例對(duì)本發(fā)明作進(jìn)一步詳細(xì)的說(shuō)明。
以下實(shí)施例僅用于說(shuō)明本發(fā)明而不用于限制本發(fā)明的范圍。實(shí)施例中未注明具體條件的實(shí)驗(yàn)方法，通常按照常規(guī)條件，例如Sambrook等人，分子克隆實(shí)驗(yàn)室手冊(cè)(New YorkCold Spring Harbor Laboratory Press，1989)中所述的條件，或按照制造廠商所建議的條件。
實(shí)施例1日本血吸蟲(chóng)雌性成蟲(chóng)特異蛋白的生物信息學(xué)分析。
1.日本血吸蟲(chóng)雌性成蟲(chóng)特異蛋白的篩選利用表達(dá)序列標(biāo)簽(EST)的數(shù)據(jù)，分析特異表達(dá)的蛋白。雌蟲(chóng)相對(duì)于雄蟲(chóng)特異表達(dá)的基因1)其差異顯著性必須滿(mǎn)足p＜0.05，這個(gè)顯著性分析用網(wǎng)上專(zhuān)門(mén)的軟件工具計(jì)算獲得；2)每個(gè)基因，其EST數(shù)量必須大于等于10條；3)雌蟲(chóng)的EST數(shù)量必須是雄蟲(chóng)數(shù)量的3倍或3倍以上。經(jīng)過(guò)這3個(gè)標(biāo)準(zhǔn)的篩選，得到了雌性成蟲(chóng)特異表達(dá)基因。
2.同源性和相似性分析日本血吸蟲(chóng)雌性成蟲(chóng)特異蛋白序列對(duì)GenBank(http://www.ncbi.nlm.nih.gov/)的非冗余數(shù)據(jù)庫(kù)(Non-redundant，NR)做BLASTP比對(duì)；日本血吸蟲(chóng)雌性成蟲(chóng)特異蛋白序列對(duì)Swissprot數(shù)據(jù)庫(kù)做BLASTP同源性比對(duì)。根據(jù)比對(duì)結(jié)果得到的與已知其他物種或已知的血吸蟲(chóng)蛋白的序列相似性，做出日本血吸蟲(chóng)雌性成蟲(chóng)特異蛋白的功能注釋。
實(shí)施例2RT-PCR實(shí)驗(yàn)驗(yàn)證日本血吸蟲(chóng)雌性成蟲(chóng)特異蛋白的表達(dá)情況。
1.樣品準(zhǔn)備準(zhǔn)備日本血吸蟲(chóng)的成年雌蟲(chóng)和雄蟲(chóng)，以抽提其RNA。
2.總RNA抽提試劑盒抽提RNA試劑采用TRIzol reagent(GIBCO/BRL)，該試劑基于酸性酚一步抽提法生產(chǎn)。用于抽提RNA所用器皿和水均要進(jìn)行無(wú)RNA酶處理，以保證實(shí)驗(yàn)中無(wú)RNA酶環(huán)境。
3.RNA抽提步驟將碾杵和勻漿器等器皿在200℃干烤4h，去除RNA酶，冷卻；加入液氮中預(yù)冷，將組織從液氮中迅速取出，碾成粉末；用刮匙將組織放入預(yù)先加入TRIzol試劑勻漿器中，勻漿數(shù)分鐘；將勻漿后液體轉(zhuǎn)入無(wú)RNA酶離管中，加入氯仿后，4℃離分層；將上層水相轉(zhuǎn)入一無(wú)RNA酶離管中，加入氯仿后，4℃離分層；將上層水相轉(zhuǎn)入一無(wú)RNA酶離管中，加異丙醇，4℃離沉淀RNA；用75％乙醇洗滌沉淀2次；用無(wú)RNA酶去離子水溶解沉淀。抽提RNA質(zhì)量鑒定紫外分光光度計(jì)測(cè)定260/20比值(比值均在1.7~2.0)；并在MOPS甲醛變性膠中觀察有無(wú)降解。
4.RT-PCR反應(yīng)用半定量反轉(zhuǎn)錄多聚酶鏈?zhǔn)椒磻?yīng)(RT-PCR)來(lái)驗(yàn)證日本血吸蟲(chóng)雌性成蟲(chóng)中一些基因的表達(dá)豐度差異。采用不同的循環(huán)數(shù)，分別是21、24、27和30個(gè)反應(yīng)循環(huán)。每個(gè)PCR重復(fù)3次。實(shí)驗(yàn)結(jié)果見(jiàn)圖1和圖2所示。
序列表<110>上海人類(lèi)基因組研究中心<120>血吸蟲(chóng)雌蟲(chóng)特異蛋白基因、蛋白及其應(yīng)用<130>NP-10282<160>46<170>PatentIn version 3.3<210>1<211>773<212>DNA<213>日本血吸蟲(chóng)(Schistosoma japonicum)<400>1acaattaacc tcgtcgtgaa atttcaagac gattacaacg ttgctgatta agcatggcta60atacaggaaa aatatctctc aaacggaagt ttgttaaagc cggagtgttt aaagctgagt120tagacgagtt ccttaaacgt gaattagctg atgacggtta tagtggtttc caggttcgtg180aaagtcgtaa atacacagag gtcatcatac tagctacgaa aactcaaagt gttcttgggg240aaggctcacg ccgtattcga gagctcacct cggttttgca aaaacgtttc ggatttcctg300atggaaccct aggactgttt gcagaaaaga tgactttccg aggcttgagt gcaatcgcac360aagcagaatc acttagatac aaactcacat atggtttacc tgtgcggaaa gcgtgttatg420gtattcttcg cttcataatg gaatctggtg ctagaggggc tgaagtcatt gtctccggta480aaatcaaggg ccaaagagca aaagctatga agttttgtga cggtcttatg attcattctg540gagacccagt gaactactac gtcgacaagg ctgttcgcca tgtccaactc ccacaaggta600tactaggaat caaggtgaag attatgatgg atcacgatag ttccggcaaa aagggtccac660ggaaaccttt gcctgataca gttacaattt tggctgctaa agaagaagtc ttaccgactg720ccccgtattt tgaaaacaaa atgtaaatac ttttgcaata aagtttgtcg tgg 773<210>2<211>805<212>DNA<213>日本血吸蟲(chóng)(Schistosoma japonicum)<400>2gcatttgttc gctctcgtga tgaagcttaa tatagcgttt ccgtccaatg gaactcaaaa60gtgtatcgac gtcgatgatg aacttcgcct tcgcccattt tttgataaac gtatatctca120tgagctccca gttgactttt tgggtgatga atggaaagga tatatagtgc gcataactgg180tgggaatgat aaacaaggct ttccaatgaa acaaggagtt ctaactaatg gccgtgtcag240attactgtta ggcaaaggga gctcttgcta ccgtcctaga cgtaagggag agagacgacg300caaaagcgta cgaggttgca tagtcgataa taatctaagt gtccttaatt tggttgttgt360
acgtaaagga gaaaatgata tccctggttt gactgatatc accattcccc gacgcctcgg420acctaaaaga gcttcgaaaa tcaggaaatt gtttaatttg tccaagaagg acaatgtatg480ccagtttgtc gtacgaaaac ctgttccagc taaagaagga aaaaaacagc gtttcaaggc540ccctaaaatc caacgtttaa caacaccttt acgattgcag cgtaagcgtc ataggcatgc600gctgaagcgc cagcgcatgg aaacccgcaa gaaaacgcaa gaggagtacg ccaaattgct660tgctcaacgc cggaaacaag cccgcgacga gcgtgcagag cgaaaacgtt cacggagtca720ttctttacgt gaatccaaag gacaaactgc caaaaatgca tgataaaatt ttttaattgc780aaataaacct atggatgagt ctttc 805<210>3<211>678<212>DNA<213>日本血吸蟲(chóng)(Schistosoma japonicum)<400>3gcgtgtggtc gattaaaagg atgtcgttgt cgaagtatct tgctcctcct ctaacttccc60taccactttg ccttcctcgc tattacggtg gggtcatctt ggtgggcgta ggtgcatttg120gattgaacgc ttacttcgta cgacgagtta tgcaagctcg caaagaacat aacgttgagt180tacccataat gtaccatcct acggacaagt tatttaactg tatccagaga ggtcatcaaa240attatctaga agttcttccc tactttctta tggctctgtt tgttggtggt ttacgctacc300ctcggacata tactgcatgc ggcgttatat tccttcttgg ccgtcttatc tattttcagg360gatattccac cggagatcca gggaagcgtt ataaaggttc tatttcaatg atcggtggac420tcccgatgat tctgggatta ctagttttcg gtgttcaaca cctggtagcc agtgtaagat480gtaccgcttg tactttacgc agataaaaga aactggattg catggaagaa tattcataaa540tacttgatat tctaatctaa cagtagtagg cattgctatt ttatcacatg tatgcattta600gaataaaatg gtataataca gcgtgtttat ctagctgaat tcactttaat attaattctc660aactaaagtg tgtaactt 678<210>4<211>724<212>DNA<213>日本血吸蟲(chóng)(Schistosoma japonicum)<400>4catgtcgcgg gcttttcaga ttttcgagag taatgatcac tggaagttcc gcaatgtctt60catctagtag atctggtagt tcaatatatg atttcactgc taaagatatt gatggcgtcg120atgtatcctt aagtaaatat cgtggttatg tttgtctcat tgtcaacgtt gcttgcaagt180gaggcgcaac ggacaaaaat tatcgccaac ttcagaagat gcacactcaa ctagttggtg240atggtctccg catattagca ttcccctgca atcaattcct tggacaagaa ccctggcctg300aagcagaaat aaaaaaattc gtgtctgaaa cgtatggtgt tcagtttgat atgttctcaa360aaataaacgt caatggccct aacgctcacc ctctttacga atttctcaaa agtagactag420gtggtgcctt aatgagtgct ataaaatgga atttctcaaa gtttctcata gatcgtaaag480gtcagccagt gaagagatat tctcctacaa ctgctccaaa tgaaatcttg gatgatatca540tggagctatt gaaacagaag tgatctaagg aatttttgag ctttgaacaa attctcgcta600
tgtgacgata gcaatctcaa gtgtttactg attgctatct gataaaatca gttttgtgtg660cacctgattg tagaattttg ttcattttag ctcatatttt cctcttgaaa atacttctac720gaat 724<210>5<211>758<212>DNA<213>日本血吸蟲(chóng)(Schistosoma japonicum)<400>5tagtttcaag ccttactggt ctctcctgaa gcagtttttg gaccaaattt ctcaggttca60tctgaatatt gcgcaggaac gacaaaatct ctccagagat gtgcaaaaat acttagagga120gcagcacaag cgacaaaagc taatcaagga tacagaagca tccacacagg aagttgttca180tgccttccag gtaacttccg ttcaacttca gaaagcaaag gaggcatacc actctcgtta240taatgaatat gagcgtgcta tgcgcttgga atctggttct aaccgcgatc aggaaaaact300tgaagtcaag cttaagaaag ctcaggatga atacaaatat tcagtcgaaa aatataataa360cttacgcaat caatttgtca gcaaaatgca catttcctgt gctcagtttg aggaaataga420agtttcacat ctagaacaga tgagagaatt tctccatcga tatgcatcgg tatggtccgt480cacaggcgat gctttagaaa aattgcacaa tcagtttgat catgatttag ctgagttaac540tactgataga ttactgaata ttcttgtcag cgaaaagggt actgggacta tggaaccaca600gagtgtacaa ttcgaagatc cagaagttgc cgctgcattt tcttcaaatg gaaatgaacg660cgctaacgga ttaacaggaa atagtgttcg ggttttcgac gcttttctta atcgcgaagg720cagagctgcc tctgcacttg caattgtagg gtctagta758<210>6<211>857<212>DNA<213>日本血吸蟲(chóng)(Schistosoma japonicum)<400>6catctgggct tgtaacttcg tagcgtgatt gttttctcgt gtgctagtta cgggcgtgaa60ttatttatgg ctgacgaaaa gaagacttct agtgacaagt cagcagttgc taatagacct120aaaaagccgg aaggtcttcg tctcaaggaa aagattaggc gccgtaagtt agttagtgct180gctcgtaaaa aagtgatgct tcgacgtact catattgtgt tgaagtctct tatgaaaaaa240cgtgctgaaa aatacatgca tgagtatcga aaaatggcaa agcgagaaat aacgtcacgg300cgtgaggcaa agaagacagg aaacttttat gttccggctg agccaaaact tgcttttgtt360gtgcgcattc gtggtataaa tggtgttcat ccaaagccac gtaaaactat gcaactgttc420cgcctaagac aaattaataa cggcatgttt gtacgattaa acaaggccac tcttaatatg480ctgcgaataa ttgatcccta catcgcatgg ggttatccta gtcttaaact tgttcgtcag540ctgatttata aacgcggatt ttgtaaatat catggttcac gactcccttt aacaaacgat600ctaatccgac gtaagcttgg acatcttggc ttaatatgtg tggaggatct tgtgcatgaa660atatacacgg ttgggccgaa cttcaaacaa gcagttggtt tcctgtggac atttaaatta720aataatccca gtggtggttt ccgcaagaaa ggaaaacatt ttgttgaagg tggagacttt780ggaaatcgag aaacctatat aaatcagtta ctaagagcta tgatttaaat aaataagctt840
tcactc ggta actccgg 857<210>7<211>1005<212>DNA<213>日本血吸蟲(chóng)(Schistosoma japonicum)<400>7ctcatttctg agtcatattt acggccatcg atattatgga acgtctactt cgacttggtg60ctgatggatt agctgcttta ggccagtcta atgcccaacc aactgacggt cctgtaccag120atactgctga gcaggtgtat attagtagtt tagcactact aaagatgctt aaacatggga180gagctggagt accaatggag gttatgggac taatgttggg tgaattcgtt gatgactaca240ccgtaacggt tgtggatgtg ttcgctatgc cccagtctgg caccggtgtg agtgttgagg300ctgttgaccc tgtttttcaa gctaaaatgt tggatatgct caaacaaacg ggacgtcctg360agatggttgt cggatggtat cactcacatc ctggatttgg ttgctggtta tcgggtgtag420atatgaatac acaacaaagt tttgaagccc tatctgatcg tgcagtagct gttgttgttg480atcctataca gagtgtgaaa ggcaaagttg tcattgatgc atttcgttta atcaacccaa540gtcttgttat cgctaaccaa gagccaagac agacaacatc taatgttggc catcttagta600aaccaacaat ccaagcactt atccacggtc ttaacagaca ctattactct ttgcccatta660attaccgcaa aaacaagtgg gaaatcaaga tgttgatgaa tttgaacaaa cgtacatgga720gggacggatt aacattggag gattataatg ctcattgctc agcaaatcac aaaacacttc780agactatgct tgatttagtc aagtcatacc ccaaatctct tgaggacgaa gaaaagatga840ctcctgagca gttggccatc aaaaatgtgg gaaaaatgga tccaaagcga catttgggag900agcacgtaga tgaactgatg acttccaaca tcactcagtt tttgggagct atgttgcatt960cagttgtttt caccatagac agttgttaaa taaatttttt tacag1005<210>8<211>1908<212>DNA<213>日本血吸蟲(chóng)(Schistosoma japonicum)<400>8atcaaattaa gtatcaaaga attaattaat aatatttaac gatgccgtta acctcacgtt60cacctcatga cagtaaaaca ctacttgcta aatattacga tttaccgcaa cctgatgata120aagttcaagt tatgtatgta tggatagatg gaagtggaga gaatttgcgc tgtaaaacaa180tgactttacc tgatgaacca caagctccag aagattgtcc catgtggaat ttcgacggaa240gtagtacagg tcaagctgaa ggttccaatt ctgatgttta tctaaaaccg tgtgcaatgt300tccgtgatcc atttcgtggt ggtaaaaata aattagtctt atgtgaaacc tttaattatg360ataaaaaacc acatgtttca aaccatcgtt atctttgtaa tttagtcatg gagaaagcaa420aaattcatga accatggttt ggtattgaac aagaatatgc attactagat attgatggtt480atccattaca atggcctaaa aatggtttcc ctccacctca gggaccttat tattgttctg540tcggggctgg taaagcactt ggtcgcgata taccagaagc tctatacaga gcttgtatgt600atgcaggagt caaaatttgt ggtagcaatg cagaagttat gccatcacag tgggaatttc660aagttggacc atgtgaaggt gtttcagccg gtgatcattt atggatggct agatttatat720
tacatcgtgt agctgaagat ttcggtgtta ttgtcacact ggatcctaaa ccgatgcctg780ggaattggaa tggttctgga gctcatacaa actattctac tttggctatg agaaatccaa840aatcaggctt acaagcaata gaggatgctg ttgagaaact gtcgcataaa cacaccgagc900atattaaatg ttatgatcct aaacatggtt tagacaatca aagacgtttg actggtagtc960atgaaacaag cagtattaat gatttttcat cgggtgttgc aaatagaggc agtagtatac1020gtattccaag acaagttgct gaaaatggtt gcggttattc tggaagatag acgcccgtca1080tctaattgtg atccatacgt agtcactaga atgttggtag aaacaacatg tttttaacca1140ctatcattat tatttttttt caaattgaca aaaaaatttc gaaaaaatat caaagaaatt1200aatatttaca aaattactct tacattacat aatctctgtg tctcatgtaa tcaatttaac1260ttaaacacgt cattctatta atgattacaa taaattagca gatttgttct aataattctg1320gtcatcatta tcgttattgt cattgtcatc ttcattacag tttttttcat cgttaacagt1380catcattgtc ataagaatta ttaacagcaa caacaacatc agaatcagca tcactttcat1440gttgtccacc gatgattcaa catagttttt tttccatgta tttttagcac ttttttattg1500atatatattc atagattact taacaaccta attgattgtc ttttgatcac tataagtcat1560tacatcattt aaactatttc tatacaaatc aaaatattga atgttagatg ttctttttgt1620tatacttcaa acatcaagtt ttctacttct tgttctatac tactgatttt actcaaaatt1680ttcatttgtg attttcattt aagtaatcag gatttttgtt gttacttatt tttaaaattg1740tgaataaaaa tattagaaac aaaaaaaaaa gaatattctt cagttgagag catttctcct1800ttgattacat aataaattat agtcttgaat atgattgatt gcgaatgaaa agtgattgct1860aaacacagag tactgaaaaa ttattattat tattatattc taatttgt 1908<210>9<211>1195<212>DNA<213>日本血吸蟲(chóng)(Schistosoma japonicum)<400>9cggggggcac gttttgcagt tcacgttaac atggctgtta caaggcctat tgtcaatgtt60tatggagtga acgggacgtc aacgggagcc accataaaga tgccggcggt atttactact120ccgcttcgct tggatctact gaatttcgtt catgatcaga tgcgaaaaaa ctcaagacaa180ccatatgcgg ttagtacaaa agctggccat caaacttctg ctgagtcctg gggtactggg240agagcagttg ctcgtatacc tagagtacgt ggtggcggta cacaccggtc aggccaaggt300gcttacggta atatgtgtcg tggtggtcat atgtatgctc caactaagac gatgagacga360tggcacagaa aaataaatat tcagcagcgg cgttatgcca tttgttctgc aatcgctgcc420actggggtac cgtcgttggt catggctcgc ggacaccgtg ttgatcgtgt tccggaactt480ccacttgttt tatctaaaga ggttgaatcg cttaaaaaga caaaagatgc tgtaaaggta540ctcaaggcaa tacgggcttg gccggatatt caaaaggttt acaactcaca acgatttcgt600gcaggtaaag gtaaaatgag aaatcgaagg cgaattcaaa aacgtggccc aataataatt660tataagaagg atgatgggat atcgcgtgct tttagaaata ttccaggtgt caccttgatt720aacgtgagag ctttaaatct ccttaagctg gctccgggtg gtcatatggg acgtttagct780atttggtcag atgcagcatt tcggcagtta gacaagcttt atggcacacg aaaaacagga840agtgtgaaaa agtcgggttt ccatctccct atagctaaaa tgaccaattc ggaccttggt900agacttttgc attgccgtgc agtttcctca gtcgtaaaat ctagacgtat gcatttgcaa960cgtccaggtc caaaagtcaa gaaaaatcca ttaaaaaatc aacaagctct actgaagttg1020
aatccttttg cactcggaga aaaacacttc aaaatggctc ttgcgcgtgc cattataaaa1080ggaaagctca agcgaaaagc agctaacaaa gttgacgaaa caccttcaaa gaaggctaaa1140actgtaaaag ttaaataata tctttaaata tagttgtgat aatggcatta aaaaa 1195<210>10<211>1437<212>DNA<213>日本血吸蟲(chóng)(Schistosoma japonicum)<400>10taagaaaatc caaaccttct gttgagtata ttgaatcagt cgatggctgc aaacagttta60tcgataatgc tacaatagcc gtccttggat ttatcaagga tacggactct ctagatttaa120cagatttcga aaaagtggct gatgaactgg acgatgcgga ttttgcagta gccaactcca180gtgatattct gaatgagtat ggtattacac agacaccaag aattgtcttg ttcaaaaatt240ttgacgaaaa tcgtgttgat tatactggaa aaacattgga aaacctgaag cacttcgtcc300aggtggaaag tgttccactg gtctcagaat tttctcagaa gaccgcaggg gttgtgttcg360gaagtccggt tcaaaaacat atcgtattct tcctttccaa atctgcggac cacttagatt420atgttaacag actgaccgaa gttgcgaagc agttcaaaag caagcttcac gttatatacg480tcgatgttga tgttgaaaac aatttacgtg ttttggaatt tttcggactt tcaaaaaatg540atgccccaac ttaccgaatt atcgagttgg gtgaagaaac aactaaatac aaaccggaca600ccgatgatta ttctgtttct ggcatgtccg attttgtaca aagagcactt gacggaaaag660ttaaaccttt ccttatgagt gaagaaattc caactgatca aacaggtgct gttagggtcc720ttgtcgggaa aaattacaat gatgttgtta gggacagatc aaaggatgtc tttgttaaat780tgtatgctcc ttggtgcggt cattgcaagg ctcttgctcc agtctggaac gaattgggtg840aagcttttaa agacgcagat gtagtgattg caaaaatgga tgctacagtc aatgaagtag900aagatctcag agttactagt tttccaactc tgaagtttta tcccaaaaat tcggatgagg960ttatcgatta tacgggcgat agatcttttg aagctttgaa gaagtttgtg gagtcaggtg1020gcaaatctag tgaaacaact aaagaagagg atcaaataaa ggatgaactc tgaacttact1080ttgtagaacg atctactttg ctttttctca ctgctctcac ttcccttggc tttttgaatc1140ttttttctcc attacctgtc aataaaaatt gggcaatagt ttaacttgcc cattaactag1200ttatttgtag ccatatgtat ttgtttttaa ggcgctatct tacagtgcat ttccatgtga1260atgttcctca agttttttta taacctacgt gcaccataaa tgttttttct ttctaaatat1320ttttaaattg tgttaaatta aattcatatt cacttgtaaa tttaactttt ggaactatgg1380aaacatttgt ttgcatcgaa tgtacaagaa agtttaccaa aaaattgtgt tgtttag 1437<210>11<211>708<212>DNA<213>日本血吸蟲(chóng)(Schistosoma japonicum)<400>11ttgcgtctcg taaattgctt gctaatgttt aacccgtatg catctaatgg gggaaccgtt60atggggattt caggcaatga ttttgccctc atagcctctg acactcgcct atgtgatggt120gatttcgtta tactctcaag gaactgtccc agacttttta aattgggtcg tggtaatgtt180
cttggatgtg ccggatttca tggggacgtc ttaacattaa cgaaactgtt agaaaacaaa240gtgaaaactt atcgttacga ccatggtaaa gatatttcaa cgaaagcgtt ggctggcctg300ttatctgtgt ctctatataa cagacgtttt ttcccatttt acgtaagcaa tatactagtt360ggtttggatg gagatcgcgg tgttttatac agctacgatc cggtaggatc atatgaagcc420gaaggttacc gcgcatctgg cagttctgca gcgatcttgc aaccattttt ggatagcacg480gtcggaaaca agacctcgaa ataccacgta tcacgtttaa acaaagaaac agcagttaaa540cttactcatg atatatttat tagcgctgca gaaagagaca cgcagtgtgg tgacggtgtt600gtaatttaca cagttactaa agatggttta agtgagacct cttatccatt gagacgtgac660tgagactatt ttggaacttt ttcatacata aagtactgtt tgtaaaat 708<210>12<211>461<212>DNA<213>日本血吸蟲(chóng)(Schistosoma japonicum)<400>12ctaaaacgtg gtgctcctcc gtggctaagg cttattgcag ctcacgtttt taatcatgtc60taaaagcgag ttagcttgtg tgtacgctgc attaatgtta gctgatgatg acatcgacgt120gacagccgac aaaatcaata ccattttgaa ggctgcaaat atcaagttcg tcgaatcata180cctccccaat cttttcgcga catctttaaa tggtaagaat gtgaaggatc tactaatgtc240catgggttca cctgcacctt cagctgcagt aacgtcggca gttccaacag cagcgtcagc300taccagtgct gctgctgaga agccaaagga agctgtgaag gaggaaaaga aggttgtctc360ggatgacgat tctgatgagt ccatcggctt tggcttgttt gattaattgc actatatgta420cagtataaat tggatgcgaa tgtgttcaaa aaaaaaaaaa a461<210>13<211>1164<212>DNA<213>日本血吸蟲(chóng)(Schistosoma japonicum)<400>13tcccaattaa ttccacatag aaatatgcgc atatatacct tcaaccaaca ggctataaaa60acagcaaata tttctattat gtacaaaatg aattaatatt cagggacaaa gtcgatgacg120cattttaagg gaaatgataa ataaaatacc tatttgcttc tactggaaag gtgagacata180gaaaagcacg gttttacatc aatgtactgt taacagacct caacagctaa caataataat240aattattatt attattttca caatcaactt acgtcccaaa tacgtcttca aatgagataa300aggctaactc attgattcgc cgaagtaaat gggtgcaacg gacaaatttt accaatagta360ttgagattta ggatgcatat aacttaccaa tttcacccta aaagctgaaa tcattatatc420caatttttct ctcaccgtaa tacgtcataa tctatttgga tgctcaagtt atgcattaaa480aactttaatg aaatgtggtg aaatatagta gaattgaatg gctgttgaca gtctctcaaa540cttaacaggc ataactatga atgaaaatca ctgtactcat tgaaaacaaa aaggtattcg600aaccatcctt cagtaaactg acagttgtct cttcaggagc agcaggctcg aactcagaga660tgataccatt gcacgcaggt gacaagttgc tgttctcatc tgacttaacc tggtggtggt720tgtggtggtg gacccgactt ttatggcaaa ggatatgagg actcgtatgg tggcgacagc780
tacggaaatg actactatgg cgattcgaat ggaaggaaga atggtcatgg taaaggaggc840aagggtggca atggtggtgg cggcggcaaa ggtggtggta agggtggtgg caatggtaag900ggtaacggga aaggaggtgg tggtaagaat ggtggtggca aaggtggtaa cggaggcaaa960ggaggcagtt atgcaccctc ctattattga gttcatcact catcaattca tgcccatcta1020tatcactcca acattcattt gtgtcatact gctctcattc ctctcatcat acaacacaca1080ccactcaatt catctcactg taatatcgta atgtgtgaat tccagataaa taaatattat1140ccattctaaa aaaaaaaaaa aaaa 1164<210>14<211>864<212>DNA<213>日本血吸蟲(chóng)(Schistosoma japonicum)<400>14aagtaataac catttattat agagaaatcg tcaatattgt ataatacaag cagatatccc60gtaaaggagg cgattaggca atttatcttc taagaattct tttcgtataa ctttcaagct120tcgtaaacac tcgcatagca aagctttcgc caaacttttg aaaaattaat cgtccatttc180tttgtgtatt tatcattttc tgcagtagct acttcgatgg tttaaaacat ggagtcgcgc240tgtttattgg aacacacaag ttgtaactgt gcatccagcg tctacagtgt ctcagcataa300tatcaatagt aaaattactg tctcttaaat gtgctagcaa catgtgccga catggataac360ggcacttgat gtacgttgga caggtgcact cacccgaagt cagatccact tcatatgatt420tcgatccttg caatatcatg aaatactttg ttagtgttgt aatcatagcg acaattggtg480ttgctttatc cttcaacatt cagaatactg atatgcaaaa cacaaaagat atgggtgatt540atcaatttga cgataatgct acctgttcaa tgtgcattaa ttatgtttcg cattggcaaa600aacatttgaa ttcaactaaa gttaaagagc ttcttcgaac ttacgctgaa ataacctgtg660acttctcatt gttattcaga tatcggtgta tacaagtgat gaacaaatac tataatgcat720tcatttcttt catagaaaat catgatgctg aaacaatatg caagtttact ccattctgtt780aaagataaaa catagaagac ttggatatga acttttacaa gattgttttt agtttatttt840aaataaaata atatttattt catc 864<210>15<211>1189<212>DNA<213>日本血吸蟲(chóng)(Schistosoma japonicum)<400>15tagtcggtct atgattgcct tgtggacgat tttgttaccg ttaatttatt atgtggatgc60tgaaccatac agagattgtg gatctaaaat tggatcatta atttcactga ctgtaacccc120ttgcgataaa gtaccatgtg ctttgtataa aggtcaaaac tctacaatta caattgaatt180cagtacaaaa gagactgtaa aaaatggtca catttctgta catggaataa ttgctcatgt240accagtacca tttgtacttg ataagtcaga tttatgtgaa tttgtcagtc caacatgtcc300attaattcca tctatacaga aatatacaca cacatattct ctgcatgtca ataaaatgta360tccttcgatt tccttaactg ttcgttggga gctacaagat agttcaaatg aagatgtgat420ttgtgttgaa ttccctgttc aacttatcta gcatcatagt atattcattg atttagtgtg480
actaaccatt catcttgttc tctttcctgt cccctcagta aataattatt tcgaacaaaa540ccaaggaaaa cctactactc attttaatca ctaaaaacat aaaacaatta cttttgacga600ttcaatcatg tgattatttc tactatcatt cctaatgcat ttcatctgat tatttagtaa660aataaagtac tttcaatcat tttaaataca caaacaattt taatcgatgt ggttttgtgg720agatatatcg atgctgagtt cagttgattg taattgttgc agtttgattt caattgagta780ccattgacaa ccgtggagca ctagactact gcttcgtcct agtatagact cctcagacgt840gcgctgtcag atcaagtctg tcgacgaata cctttccatt agcaccactg ggtcggccaa900tgatagaatt ctcactttca acaatttatg atatagtgca gcgatcagtt aattctgttg960gtaataactg tctcactccc gatatatctg aactccacta gttacaactt ttcactagaa1020cttcgagaaa atcttaaagt taatcaataa tgagctgatt attgtagttg aaatgaggtg1080gtatgaagtt tagttgatta tatactgcaa tgaaacggct ttccactgct ccctgtttat1140cagtggtacc caaccgacat ccatcaacta aataaacagt ttgtttggc1189<210>16<211>1344<212>DNA<213>日本血吸蟲(chóng)(Schistosoma japonicum)<400>16attgtaaaac aattattgta aaattaggta aattgcataa tattaacata accgtaatga60attgttcatt tttcaatata tttcttattt tattgatttc tattgcttat acaaaatgta120tgccacattt tattactggt aagatataca taccaatatc atttggtgct tttgcaactg180tggattatcc aattataaga ttttggactg gacgtttcta tagactatgt gggagatcat240attactgtga ttattataat aatgatgaat atgatgatga ttattcttat tctgattata300attcaaaatt ggcaacacct tatcaacaaa agttattacc accatcatca tcatcttata360acaaatatga aaataattat aattatgaag cgttggacaa taaacaacca aaggaaaatt420catattctta tgctgaaaat gttaaaacag ataaaatttc atctatgtat caaaaatatg480atgaaaatta taagaaagat gattataaaa cgcctattta tgctgatact aaatacacta540aaccaactta taatagaatt atcaaaacga aatctgaaga tgtatataat aacaaatata600atgacaaaac ttttgattct acaagtcatt attataagaa atatttaact agagaaccta660cgtacaagta taataaacct attgtgcata aatctaagaa gtcagaatat gatactcaac720aatctaaggt attaaaatct gcctatacta atgataaaac taagaaacat cacaataaac780aagacgctta tcaaccgagt gtagataata atcatcctat cagtgaacac cgatccggat840ttttaaatcc tgatcatatt tatggtgatg aaattatgcg aacaaagtac tatttcgatc900aattcactga tgacgtagtt agacaaattt ctaaggataa caataaatgg aatgaacaaa960catatcaaca acctaaatat attcaaaaaa agtcacatga taattataaa actcgatata1020ggaagaaaat ggataaaaaa ggtgaaaaga taaaaagcaa tgaatataaa aagacaaaac1080ataataacta tgttaataaa gtacctgatt ataataacaa gttgataggt agaaaattcc1140caaaaaataa aatggccgaa tcaaaatata gagagactgt ttatcaaaag aaggtaaaag1200aacaaactaa accacgggat ggagattatg ctgccaaggg ctacgcaaaa gaataatgta1260ttgaatagaa tagtaatata tacatatata tatatatgat ttaattctaa ataaagtatt1320gtaacaaaaa aaaaaaaaaa aaaa 1344<210>17
<211>692<212>DNA<213>日本血吸蟲(chóng)(Schistosoma japonicum)<400>17tgacaacata ctcctattta cttgtatcat tcattcttct tgatagttat tgttccggtt60atggatatta ttactcaagt cattatcgta aacatttcga tccggctatt gcctcattta120caaaagaacc aaatataggt tctgtatggt ttacacaaca tggaaattac atgtacctta180acggaagtgt ttcgggactt ccacgtaatt caaatctagg tgtacacatt catcgatacg240ggggacttgg aaacatgtgt cttgaagctg gacctcattt taatccattc aacatgcgtc300acggaggccg tttcggaaag cctagacatg ctggtgattt aggtaatatt cgagttgacc360gatatggtgt tatggtattc gatatttatg taacaataga aaaacttgga ccatatgacg420gatttattgg gcgatcactc gtgatacatg ccaatatgga cgacctcgga agaaatccag480atgaaggcag tcgaacaact ggcaattctg gtccgagatt agcttgtgct acaataggat540atcgtggacc ataaagtaaa tttcgtttaa taaatcgaat tttacctagt tccattaaga600ataatacaat aaatttaatt gaataaaaaa aaaaaaaaaa aaaaaaaaaa aaaataaatt660taaaaaaaac ttgagttttt tttttttttt tt 692<210>18<211>989<212>DNA<213>日本血吸蟲(chóng)(Schistosoma japonicum)<400>18gttattcaat agtattagta ttttacatta acttcgttac ttccgttttt acatattatg60atgattacga gtatggaatg ataggtgaag ctggcagaga tgcagaagtt gctttacagt120atacagtggg aaaatttgat gggaaaaaat atggagaaaa ttatgaatat ttacaagata180attatggtaa atacggtgat tatggtgatt atggatatga ttcatattat ggggatgaag240gatacgatta taatcacgtt actttagctg acagagcaat gcacaaatta aaagacatga300aacgtcgtaa acatcattat ctatcaaaag attatgatga aggatatgga tatggatatg360gaccagaagt agaagaatat aaagaacctg gatataatgg gaataattat cctggacgta420aaccaactat gttattgcca ccaccaacca aaccaccttt acaaagtcag aatttcagag480cagaatagtt cagaatattc aaactataaa agaatcgaaa tgtaaatgaa tcaaatgatt540taataataat tcaatataat tctgttggtt aatatgtatt gaacaatcta tttcaggaaa600atagattatc atattcctat ctttaagtgt tcaatacatt tcactgattt tctttgttaa660cttttctgtc ataccaaatg tatctcatac ttacatttgt atttggtata atttattcag720aaaactgtca gtaaaaaaaa acaaaaaatg aaaaaacatt caaaatgcaa caaaatgaat780tcctgggttt aaaaaagctt tactaatcac tccaccctgg gtggcccctg taaaaaccag840gctcgagagc caggaatctt taccgctaga ctacttggaa gttctgtttg tggactgaag900tgttactgtc cttgtgctta tccttaaggg cggtccgaaa ataaccacca atatagctta960aaatgtatgt taccttatca tttgatttt 989<210>19<211>842
<212>DNA<213>日本血吸蟲(chóng)(Schistosoma japonicum)<400>19ttgtcagagc gtgaccatga gacttatgat tgttttcctc taagaagttc tgacactaaa60cttttggatg ctaaaaatta ccgtcttttt ctagtaaggt accaaaagac ctgttgctgc120gctcatgtgc tgatatcctt gaatacagca ataagacaaa gaaacgaaag tttactgaga180ccgtcgagtt acaaatagca ttaaagaatt atgatgtcca aaaggataaa aggttctccg240ggactgtgcg gtaagtccat gaacccccac ccagggttct aaatttgtgg ggaggctcct300gccgtccgtt tcaaatcgaa tctcatttta ggctaaagca cattccgcgt cctaaaatgt360ctgtgtgtgt gctcggggac caacagcact gcgacgaggc caaggccgca ggtattccgt420gcttggatgc tgaagacttg aaaaagttcg gaaaagaaaa gaagccactg aagaaatttg480ctaaaagtta ccacgctttc gtcgcaagtg agactattat tcgtcagttc acgagaactg540tggttccggt gctgaccaag tatggaaagt ttccagctgc tgctacacac aacgagccat600tgattaataa gattgacgag atgaaggcat cagttaagtt ccaaatgaag aaggtcctat660gcctaaacgt tgctgtaggt catgtgaaga tgactggtga agaactggca cagaatataa720acttgtcaat taatttcctt gtaacacttt tcgaagaaga attggcagaa cgtcaggtgt780ttgtacatca aaagcacgat gggtaaaccc cacaggctat attaaataaa tacagttttg840gg 842<210>20<211>1195<212>DNA<213>日本血吸蟲(chóng)(Schistosoma japonicum)<400>20attcagttat tattacattt aaatatttcg ataatcaatt taagttgagt agaaattcaa60ttttgttatt tttaaaaata aaaggcggga gaaaatgaga ttaatatgct tatgcataat120aatatatacc ccattgattt attgttttag acaaggacta tatgattatg atgatgattc180atttttcaat gacagatatg atttctctag tcatgattat tcagaaagtt atgattctga240ttatcaatca cctgataaat ctcctaatca acatcattct tatgatgaac cggattcaca300tttacatcca aacaaaaatg aatcaggcgg attgtttaat tatcacaaaa acagtagtca360tcgtcatgaa aattataaac ataatcatcg taaaagtaaa tatcatcgtt acaatgatga420ttggcatcgt tatgatgatg aattaaaaga tatcattttt ggtgacggct atcaacaagt480attttctcaa tatacagaag gtcatggatt attaggtgac gaaacttatg gttcaacaca540tagagatcca tcaaaatatg atcgctatgg aaactatatg cctgattata caacaccaaa600tggatctcct aaccctccac cttcttccga tgaaggcgaa gaagaaggag acgaagaaga660agaagaagaa gaaggagact atgggggtga agaaagtgaa acagaagttg taatgaaacc720ggctccatac gttaaatata agcaatattc accacaaccc actcctatac caaatacaca780aaatatggcc gcactgaaaa gttcaaagaa aaaaaagaaa aaaaagaaaa agtctaagaa840aaagaggtat tattatgcaa agaaatcaca gaagaaacca ggagtacatt attctgttca900agctgatgga ggaagtggag gtggcaatag ttacaagcct aagcctcatt catatagcta960tcaccatcag tattattatg atattgatgt agatgactat tccaatgatg ataacgatac1020tatggaagcg atgtaaacaa aatgagaata atataagctc atcatcttaa actaattcaa1080
accattatta caagtgaatg taaaaaattg tttgcttgtt atatccatta aatatacatt1140gtttatttat ttacttgtag ataaacatat atttaaataa aataatcatt acgac 1195<210>21<211>1237<212>DNA<213>日本血吸蟲(chóng)(Schistosoma japonicum)<400>21atgttcatcg attagtcatt taatttaaaa ttcaacatcg ttattaatta gtttgttgat60tgttgtcgtt gcagttgaat taactatcta catcgttgtg aactgctact atgatgtata120caatgaagtt gttttattat aatagtggag cgtatataac tcattttatt atgtatagct180tattcatggt gttaatttta acatctggaa atgctaatga tttgtctact gattctgtgt240ctgagtctaa tgacagtaat ttaactaaca caacttctac accaacagac tcgtcaacta300caacagagag tactgatgtt tccgtaacag agagcactag tactgatgag actaccacca360ccaatcctgt taataccact gaagttgggg ttatttcaac cacttctgaa actctgtctt420ctacatcttt aagtgcctca gaagaaacta atagtgatca gaagcaaaag aacaggtcta480taatttctag tgtaactgga atggagtctg cagatatagc accaacaaca gagtcacaac540aaacgaccac ccttcaagaa aataatactt cggtcttaca taataaaaca gatgaatatc600aaaccacagt aacttcaaca gaagataatc ctaagattat tacaactaca cctaaaatta660atggtgtatc attatttcaa gttgattaca ttgtacaatt gttaattcta ttcataattc720aatttatcta ttaacattat taaataattc atttaataaa catgttaaca ttgaatagta780agtatattta actaaaaaaa ttttgtttaa aataacatta acttatcaat ttctaaaata840ttctacttac tattcattat tattaatctc atagtttgtt taatctacag ttttttttaa900aaaaagaaaa accaacatat ttttgtttgt ttatttgttt gtttaaatta ttaaatgtac960aatttttgca ttttgttttt cttttttctc ttccatttaa caaaattatt aaattacatt1020atcaataatt aaagttaagc atattttgtt ttgtttatta tataattaaa tgattaaata1080ttgtttattt gagtaaagaa attaagtatc aatagaaaga agtttattcc tatcattatt1140cactaattat atttcattat tattattatt attaagctaa atatttattt aaatgttata1200gtttttgtaa actattttgt tattcttaat gttctca 1237<210>22<211>970<212>DNA<213>日本血吸蟲(chóng)(Schistosoma japonicum)<400>22acagtgtact gagtgcactt tatcaaatta tgtttataaa tgagtatgtt gtaatgacaa60ttgtcattcc gacgcaatat tttccatgaa ataattgaca tagagaagtt gtcagtaatg120gaaacgaaag tgatgttcat aattttgatg actgtgacaa tcatattcat caaatcttca180aaagctcaaa atatgcatca acaccctcct ttaaattcca gtagtggaag ttcattttcg240agcagtgata tgagaaacca aggaagtagc agtcatagaa atgataaaaa agattcatcg300agacatgaaa agaatggtat tgcattagag accatgtcag gaacatccac atcaagatat360cagtacagga aaatgaaaga tcttaaagcc agaggcaaat cccgatcacg tggtgttgga420
agatatggta caatgtcatc tgcatcaaca gatttcgtga ttgaagggaa gatcggtaaa480tatggtcata ggaaacctat gaaatcgaaa ttcaaaactc gtggtaagga gaagaaatat540tcaaggaaga ttagtgacaa tcgtttcgat attaaaggtg atttgtatgc acgacatcaa600catgagggta ttggtgatta tcaagcgagt ggtaattata ctactctcaa actgaaaaca660aaggagaaaa gtcgtgaatc taatcgtgac aaacagaaat ctaattttaa atcacataac720aatcaatcaa atgaaggctc aaacaatcgt tggaatatta ttcagtgaat acattattca780catatttagt tgaaaatagt ctgtttcacg atattcgggt ttcttttctg tgaacacctt840cagttttcat caatctatta atagccacca ctgacattta ttgtactaca atctacttac900tgttaatcag aaatatatac attcaactac tttaaaaaaa aaaaaaaaaa actcgagttt960tttttttttt 970<210>23<211>1212<212>DNA<213>日本血吸蟲(chóng)(Schistosoma japonicum)<400>23gaggcacccc cctctattgt ataattatct catacttact caacttatat cactgaaata60ttttgctgat tattcattta gatattttca tttaaagaat aagaagagaa catgtgcact120tttgtattga gactaacatt actactaata aattcacttg ttggtctatc ggcattaata180cttgcatcat ttggtgcaat tttagtttgg ggcaaagata caatcacaaa agaattgaat240gatgtaatcg gaccaatgct taataaaatg tatggtgaag aagtggcacg tgattttaca300gatttggcat ctactatttt agaatttaca agtccattcg gtttactctt atttatatta360ggattagtag cattggctat ttgtttattt ggtttctgtg gtacaagttg taaaaatctt420ctttgtctta gaatttacat tggattactg ttagtattgg ttgttataga gattactgca480atgagttttt attacactga tcgagaatct atattcaagc tggctaggga cattgctgaa540aattcattaa gaaattatca atcaattggt tcaaataata ctgacagcat cgtttattcc600gttatcatgc caactttaaa ctgttgtggt ttagtaaatg gatctgattt tgatcatgct660cctcgttttc aaagaaatgt gttctttaat caacaaactt acaatttgca atatccaatt720tcatgttgta agatggattc aaaatttgtt atacttgatc caacgtgtcc aggagaattc780aatgcacata attcaaatat tcatcaaggt tgttggccga aactacgacc tgttattggt840ttctattgtg gagtgattgt aatttgtgga ttggtagtaa ttttatttca actactgttg900gtcatagtga cggcagtgat actgaccact ggacgtcgtt aacaaaaagg ttatatgtta960ggatgttttg aaggtaacag atagaagagt ttcattgtct gcttcattga ataattacag1020aaatgtagta ttaaagaaaa atacagtcat cgtgtatcat ttaactattc catgttcatt1080actttctata gaattctcat agtttaactt ttatgtatct cttataattt tctattgaat1140tcataatatt tcaaataaac attgtttgtt gtaacaatta gttcataaat gaaagtttct1200ctaaggaacc aa1212<210>24<211>230<212>PRT<213>日本血吸蟲(chóng)(Schistosoma japonicum)
<400>24MANTGKISLK RKFVKAGVFK AELDEFLKRE LADDGYSGFQ VRESRKYTEV IILATKTQSV60LGEGSRRIRE LTSVLQKRFG FPDGTLGLFA EKMTFRGLSA IAQAESLRYK LTYGLPVRKA120CYGILRFIME SGARGAEVIV SGKIKGQRAK AMKFCDGLMI HSGDPVNYYV DKAVRHVQLP180QGILGIKVKI MMDHDSSGKK GPRKPLPDTV TILAAKEEVL PTAPYFENKM 230<210>25<211>247<212>PRT<213>日本血吸蟲(chóng)(Schistosoma japonicum)<400>25MKLNIAFPSN GTQKCIDVDD ELRLRPFFDK RISHELPVDF LGDEWKGYIV RITGGNDKQG60FPMKQGVLTN GRVRLLLGKG SSCYRPRRKG ERRRKSVRGC IVDNNLSVLN LVVVRKGEND120IPGLTDITIP RRLGPKRASK IRKLFNLSKK DNVCQFVVRK PVPAKEGKKQ RFKAPKIQRL180TTPLRLQRKR HRHALKRQRM ETRKKTQEEY AKLLAQRRKQ ARDERAERKR SRSHSLRESK240GQTAKNA 247<210>26<211>161<212>PRT<213>日本血吸蟲(chóng)(Schistosoma japonicum)<400>26MSLSKYLAPP LTSLPLCLPR YYGGVILVGV GAFGLNAYFV RRVMQARKEH NVELPIMYHP60TDKLFNCIQR GHQNYLEVLP YFLMALFVGG LRYPRTYTAC GVIFLLGRLI YFQGYSTGDP120GKRYKGSISM IGGLPMILGL LVFGVQHLVA SVRCTACTLR R161<210>27<211>114<212>PRT<213>日本血吸蟲(chóng)(Schistosoma japonicum)<400>27MHTQLVGDGL RILAFPCNQF LGQEPWPEAE IKKFVSETYG VQFDMFSKIN VNGPNAHPLY60EFLKSRLGGA LMSAIKWNFS KFLIDRKGQP VKRYSPTTAP NEILDDIMEL LKQK 114<210>28<211>252<212>PRT<213>日本血吸蟲(chóng)(Schistosoma japonicum)<400>28SFKPYWSLLK QFLDQISQVH LNIAQERQNL SRDVQKYLEE QHKRQKLIKD TEASTQEVVH60
AFQVTSVQLQ KAKEAYHSRY NEYERAMRLE SGSNRDQEKL EVKLKKAQDE YKYSVEKYNN120LRNQFVSKMH ISCAQFEEIE VSHLEQMREF LHRYASVWSV TGDALEKLHN QFDHDLAELT180TDRLLNILVS EKGTGTMEPQ SVQFEDPEVA AAFSSNGNER ANGLTGNSVR VFDAFLNREG240RAASALAIVG SS252<210>29<211>253<212>PRT<213>日本血吸蟲(chóng)(Schistosoma japonicum)<400>29MADEKKTSSD KSAVANRPKK PEGLRLKEKI RRRKLVSAAR KKVMLRRTHI VLKSLMKKRA60EKYMHEYRKM AKREITSRRE AKKTGNFYVP AEPKLAFVVR IRGINGVHPK PRKTMQLFRL120RQINNGMFVR LNKATLNMLR IIDPYIAWGY PSLKLVRQLI YKRGFCKYHG SRLPLTNDLI180RRKLGHLGLI CVEDLVHEIY TVGPNFKQAV GFLWTFKLNN PSGGFRKKGK HFVEGGDFGN240RETYINQLLR AMI 253<210>30<211>317<212>PRT<213>日本血吸蟲(chóng)(Schistosoma japonicum)<400>30MERLLRLGAD GLAALGQSNA QPTDGPVPDT AEQVYISSLA LLKMLKHGRA GVPMEVMGLM60LGEFVDDYTV TVVDVFAMPQ SGTGVSVEAV DPVFQAKMLD MLKQTGRPEM VVGWYHSHPG120FGCWLSGVDM NTQQSFEALS DRAVAVVVDP IQSVKGKVVI DAFRLINPSL VIANQEPRQT180TSNVGHLSKP TIQALIHGLN RHYYSLPINY RKNKWEIKML MNLNKRTWRD GLTLEDYNAH240CSANHKTLQT MLDLVKSYPK SLEDEEKMTP EQLAIKNVGK MDPKRHLGEH VDELMTSNIT300QFLGAMLHSV VFTIDSC 317<210>31<211>342<212>PRT<213>日本血吸蟲(chóng)(Schistosoma japonicum)<400>31MPLTSRSPHD SKTLLAKYYD LPQPDDKVQV MYVWIDGSGE NLRCKTMTLP DEPQAPEDCP60MWNFDGSSTG QAEGSNSDVY LKPCAMFRDP FRGGKNKLVL CETFNYDKKP HVSNHRYLCN120LVMEKAKIHE PWFGIEQEYA LLDIDGYPLQ WPKNGFPPPQ GPYYCSVGAG KALGRDIPEA180LYRACMYAGV KICGSNAEVM PSQWEFQVGP CEGVSAGDHL WMARFILHRV AEDFGVIVTL240DPKPMPGNWN GSGAHTNYST LAMRNPKSGL QAIEDAVEKL SHKHTEHIKC YDPKHGLDNQ300RRLTGSHETS SINDFSSGVA NRGSSIRIPR QVAENGCGYS GR 342<210>32
<211>375<212>PRT<213>日本血吸蟲(chóng)(Schistosoma japonicum)<400>32MAVTRPIVNV YGVNGTSTGA TIKMPAVFTT PLRLDLLNFV HDQMRKNSRQ PYAVSTKAGH60QTSAESWGTG RAVARIPRVR GGGTHRSGQG AYGNMCRGGH MYAPTKTMRR WHRKINIQQR120RYAICSAIAA TGVPSLVMAR GHRVDRVPEL PLVLSKEVES LKKTKDAVKV LKAIRAWPDI180QKVYNSQRFR AGKGKMRNRR RIQKRGPIII YKKDDGISRA FRNIPGVTLI NVRALNLLKL240APGGHMGRLA IWSDAAFRQL DKLYGTRKTG SVKKSGFHLP IAKMTNSDLG RLLHCRAVSS300VVKSRRMHLQ RPGPKVKKNP LKNQQALLKL NPFALGEKHF KMALARAIIK GKLKRKAANK360VDETPSKKAK TVKVK 375<210>33<211>356<212>PRT<213>日本血吸蟲(chóng)(Schistosoma japonicum)<400>33RKSKPSVEYI ESVDGCKQFI DNATIAVLGF IKDTDSLDLT DFEKVADELD DADFAVANSS60DILNEYGITQ TPRIVLFKNF DENRVDYTGK TLENLKHFVQ VESVPLVSEF SQKTAGVVFG120SPVQKHIVFF LSKSADHLDY VNRLTEVAKQ FKSKLHVIYV DVDVENNLRV LEFFGLSKND180APTYRIIELG EETTKYKPDT DDYSVSGMSD FVQRALDGKV KPFLMSEEIP TDQTGAVRVL240VGKNYNDVVR DRSKDVFVKL YAPWCGHCKA LAPVWNELGE AFKDADVVIA KMDATVNEVE300DLRVTSFPTL KFYPKNSDEV IDYTGDRSFE ALKKFVESGG KSSETTKEED QIKDEL356<210>34<211>212<212>PRT<213>日本血吸蟲(chóng)(Schistosoma japonicum)<400>34MFNPYASNGG TVMGISGNDF ALIASDTRLC DGDFVILSRN CPRLFKLGRG NVLGCAGFHG60DVLTLTKLLE NKVKTYRYDH GKDISTKALA GLLSVSLYNR RFFPFYVSNI LVGLDGDRGV120LYSYDPVGSY EAEGYRASGS SAAILQPFLD STVGNKTSKY HVSRLNKETA VKLTHDIFIS180AAERDTQCGD GVVIYTVTKD GLSETSYPLR RD 212<210>35<211>116<212>PRT<213>日本血吸蟲(chóng)(Schistosoma japonicum)<400>35MSKSELACVY AALMLADDDI DVTADKINTI LKAANIKFVE SYLPNLFATS LNGKNVKDLL60
MSMGSPAPSA AVTSAVPTAA SATSAAAEKP KEAVKEEKKV VSDDDSDESI GFGLFD116<210>36<211>122<212>PRT<213>日本血吸蟲(chóng)(Schistosoma japonicum)<400>36MAKDMRTRMV ATATEMTTMA IRMEGRMVMV KEARVAMVVA AAKVVVRVVA MVRVTGKEVV60VRMVVAKVVT EAKEAVMHPP IIEFITHQFM PIYITPTFIC VILLSFLSSY NTHHSIHLTV120IS 122<210>37<211>137<212>PRT<213>日本血吸蟲(chóng)(Schistosoma japonicum)<400>37MYVGQVHSPE VRSTSYDFDP CNIMKYFVSV VIIATIGVAL SFNIQNTDMQ NTKDMGDYQF60DDNATCSMCI NYVSHWQKHL NSTKVKELLR TYAEITCDFS LLFRYRCIQV MNKYYNAFIS120FIENHDAETI CKFTPFC 137<210>38<211>146<212>PRT<213>日本血吸蟲(chóng)(Schistosoma japonicum)<400>38MIALWTILLP LIYYVDAEPY RDCGSKIGSL ISLTVTPCDK VPCALYKGQN STITIEFSTK60ETVKNGHISV HGIIAHVPVP FVLDKSDLCE FVSPTCPLIP SIQKYTHTYS LHVNKMYPSI120SLTVRWELQD SSNEDVICVE FPVQLI 146<210>39<211>417<212>PRT<213>日本血吸蟲(chóng)(Schistosoma japonicum)<400>39CKTIIVKLGK LHNINITVMN CSFFNIFLIL LISIAYTKCM PHFITGKIYI PISFGAFATV60DYPIIRFWTG RFYRLCGRSY YCDYYNNDEY DDDYSYSDYN SKLATPYQQK LLPPSSSSYN120KYENNYNYEA LDNKQPKENS YSYAENVKTD KISSMYQKYD ENYKKDDYKT PIYADTKYTK180PTYNRIIKTK SEDVYNNKYN DKTFDSTSHY YKKYLTREPT YKYNKPIVHK SKKSEYDTQQ240SKVLKSAYTN DKTKKHHNKQ DAYQPSVDNN HPISEHRSGF LNPDHIYGDE IMRTKYYFDQ300
FTDDVVRQIS KDNNKWNEQT YQQPKYIQKK SHDNYKTRYR KKMDKKGEKI KSNEYKKTKH360NNYVNKVPDY NNKLIGRKFP KNKMAESKYR ETVYQKKVKE QTKPRDGDYA AKGYAKE 417<210>40<211>183<212>PRT<213>日本血吸蟲(chóng)(Schistosoma japonicum)<400>40TTYSYLLVSF ILLDSYCSGY GYYYSSHYRK HFDPAIASFT KEPNIGSVWF TQHGNYMYLN60GSVSGLPRNS NLGVHIHRYG GLGNMCLEAG PHFNPFNMRH GGRFGKPRHA GDLGNIRVDR120YGVMVFDIYV TIEKLGPYDG FIGRSLVIHA NMDDLGRNPD EGSRTTGNSG PRLACATIGY180RGP 183<210>41<211>161<212>PRT<213>日本血吸蟲(chóng)(Schistosoma japonicum)<400>41YSIVLVFYIN FVTSVFTYYD DYEYGMIGEA GRDAEVALQY TVGKFDGKKY GENYEYLQDN60YGKYGDYGDY GYDSYYGDEG YDYNHVTLAD RAMHKLKDMK RRKHHYLSKD YDEGYGYGYG120PEVEEYKEPG YNGNNYPGRK PTMLLPPPTK PPLQSQNFRA E161<210>42<211>149<212>PRT<213>日本血吸蟲(chóng)(Schistosoma japonicum)<400>42MSVCVLGDQQ HCDEAKAAGI PCLDAEDLKK FGKEKKPLKK FAKSYHAFVA SETIIRQFTR60TVVPVLTKYG KFPAAATHNE PLINKIDEMK ASVKFQMKKV LCLNVAVGHV KMTGEELAQN120INLSINFLVT LFEEELAERQ VFVHQKHDG 149<210>43<211>313<212>PRT<213>日本血吸蟲(chóng)(Schistosoma japonicum)<400>43MRLICLCIII YTPLIYCFRQ GLYDYDDDSF FNDRYDFSSH DYSESYDSDY QSPDKSPNQH60HSYDEPDSHL HPNKNESGGL FNYHKNSSHR HENYKHNHRK SKYHRYNDDW HRYDDELKDI120IFGDGYQQVF SQYTEGHGLL GDETYGSTHR DPSKYDRYGN YMPDYTTPNG SPNPPPSSDE180GEEEGDEEEE EEEGDYGGEE SETEVVMKPA PYVKYKQYSP QPTPIPNTQN MAALKSSKKK240
KKKKKKSKKK RYYYAKKSQK KPGVHYSVQA DGGSGGGNSY KPKPHSYSYH HQYYYDIDVD300DYSNDDNDTM EAM 313<210>44<211>207<212>PRT<213>日本血吸蟲(chóng)(Schistosoma japonicum)<400>44MMYTMKLFYY NSGAYITHFI MYSLFMVLIL TSGNANDLST DSVSESNDSN LTNTTSTPTD60SSTTTESTDV SVTESTSTDE TTTTNPVNTT EVGVISTTSE TLSSTSLSAS EETNSDQKQK120NRSIISSVTG MESADIAPTT ESQQTTTLQE NNTSVLHNKT DEYQTTVTST EDNPKIITTT180PKINGVSLFQ VDYIVQLLIL FIIQFIY207<210>45<211>216<212>PRT<213>日本血吸蟲(chóng)(Schistosoma japonicum)<400>45METKVMFIIL MTVTIIFIKS SKAQNMHQHP PLNSSSGSSF SSSDMRNQGS SSHRNDKKDS60SRHEKNGIAL ETMSGTSTSR YQYRKMKDLK ARGKSRSRGV GRYGTMSSAS TDFVIEGKIG120KYGHRKPMKS KFKTRGKEKK YSRKISDNRF DIKGDLYARH QHEGIGDYQA SGNYTTLKLK180TKEKSRESNR DKQKSNFKSH NNQSNEGSNN RWNIIQ 216<210>46<211>276<212>PRT<213>日本血吸蟲(chóng)(Schistosoma japonicum)<400>46MCTFVLRLTL LLINSLVGLS ALILASFGAI LVWGKDTITK ELNDVIGPML NKMYGEEVAR60DFTDLASTIL EFTSPFGLLL FILGLVALAI CLFGFCGTSC KNLLCLRIYI GLLLVLVVIE120ITAMSFYYTD RESIFKLARD IAENSLRNYQ SIGSNNTDSI VYSVIMPTLN CCGLVNGSDF180DHAPRFQRNV FFNQQTYNLQ YPISCCKMDS KFVILDPTCP GEFNAHNSNI HQGCWPKLRP240VIGFYCGVIV ICGLVVILFQ LLLVIVTAVI LTTGRR 27權(quán)利要求
1.一類(lèi)日本血吸蟲(chóng)雌性成蟲(chóng)特異蛋白，其特征在于，選自下組(a)具有SEQ ID No.24～SEQ ID No.46所示氨基酸序列的蛋白；(b)由(a)的蛋白發(fā)生一個(gè)或幾個(gè)氨基酸置換、缺失或插入而形成的具有(a)蛋白相同活性的蛋白。
2.如權(quán)利要求1所述的蛋白，其特征在于，它具有SEQ ID No.24～SEQ ID No.46所示的氨基酸序列。
3.一類(lèi)日本血吸蟲(chóng)雌性成蟲(chóng)特異蛋白基因，其特征在于，它編碼權(quán)利要求1所述的蛋白。
4.如權(quán)利要求3所述的基因，其特征在于，選自下組(a)SEQ ID No.1～SEQ ID No.23所示的核苷酸序列；(b)SEQ ID No.1～SEQ ID No.23所示的核苷酸序列中每條序列的互補(bǔ)序列；(c)與SEQ ID No.1～SEQ ID No.23所示的核苷酸序列中每條序列有至少70％同源性、且編碼相同功能蛋白質(zhì)的核苷酸序列。
5.如權(quán)利要求3或4所述的基因，其特征在于，它具有SEQ ID No.1～SEQ IDNo.23所示的核苷酸序列。
6.一類(lèi)日本血吸蟲(chóng)病防治疫苗，其特征在于，所述疫苗以權(quán)利要求1所述的蛋白為免疫原。
7.一類(lèi)能與權(quán)利要求1所述的蛋白特異性結(jié)合的抗體。
8.權(quán)利要求1所述的蛋白在制備預(yù)防和治療日本血吸蟲(chóng)病的藥物中的應(yīng)用。
全文摘要
本發(fā)明公開(kāi)了日本血吸蟲(chóng)(Schistosoma japonicum)雌性成蟲(chóng)特異蛋白及其基因序列，還公開(kāi)了與該蛋白特異結(jié)合的抗體，以及把雌性成蟲(chóng)特異蛋白作為免疫原制備所得的日本血吸蟲(chóng)病防治疫苗，此外，本發(fā)明還公開(kāi)了日本血吸蟲(chóng)雌性成蟲(chóng)特異蛋白在制備預(yù)防和治療日本血吸蟲(chóng)病的藥物中的應(yīng)用。運(yùn)用本發(fā)明的蛋白，可抑制血吸蟲(chóng)雌性成蟲(chóng)的產(chǎn)卵，從而治療血吸蟲(chóng)病、防止血吸蟲(chóng)的擴(kuò)散。
文檔編號(hào)A61K39/002GK1986805SQ200510111728
公開(kāi)日2007年6月27日 申請(qǐng)日期2005年12月20日 優(yōu)先權(quán)日2005年12月20日
發(fā)明者韓澤廣, 劉鋒, 王志勤, 胡薇, 馮正 申請(qǐng)人:上海人類(lèi)基因組研究中心, 中國(guó)疾病預(yù)防控制中心寄生蟲(chóng)病預(yù)防控制所