日本血吸虫大陆株抱雌沟蛋白的基因序列

摘要

本发明涉及生物基因领域，具体涉及日本血吸虫抱雌沟蛋白的基因序列及在大肠杆菌中的表达。本发明根据已报道的曼氏血吸虫编码抱雌沟蛋白的基因SmGcp序列和编码日本血吸虫抱雌沟蛋白保守区的cDNA序列SjGcp1设计两对引物，分别扩增出日本血吸虫抱雌沟蛋白编码序列的5’和3’区段，再根据测序结果设计编码全长引物，从虫体中抽提总RNA，用PCR克隆出该基因，克隆后经全序列测定表明该成熟蛋白cDNA编码1932个核苷酸，即643个氨基酸。该基因的成功克隆及基因序列测定为以后阐明血吸虫的生殖发育机理，以及从阻滞雌虫成熟方面防治血吸虫病打下了良好的基础。

主权项

1、一种编码为日本血吸虫抱雌沟蛋白的核苷酸序列，其特征在于该核苷酸序列及编码相应的氨基酸序列如下：gttacactgt tatactgaag tagtttaaat gaaatatgta ataataaatc aataagtaat 60tatacataac atttaattag tattactgtt tttggacaat ctataaaatt tggtaaaata 120tctgacaaag ttagttatgc caatccagat caagcagggt atcatcattg tgcattttgg 180cgtttatctg atggtcaatc aattattgct tcatgcactc cattcacatt acaacgctgt 240gggaaaaagt tagaattcga ttacaaatgt tgtgcgggat atgaaaagat tggaaaatta 300tcatatacat ttcgtgattt caagaaggat gaatgcgctc tacaagtatc cccatatgaa 360ttgtgc atg gaa tca att cgg aat cac tcc gat gta tat cca ttc gca 408 Met Glu Ser Ile Arg Asn His Ser Asp Val Tyr Pro Phe Ala 1 5 10gaa aaa atg aaa aca ctc aat gaa ctc aac aca aag aat cca gat aaa 456Glu Lys Met Lys Thr Leu Asn Glu Leu Asn Thr Lys Asn Pro Asp Lys15 20 25 30cca tac aca att ttt gca cca agt act aga aac gcc gag aaa tat aaa 504Pro Tyr Thr Ile Phe Ala Pro Ser Thr Arg Asn Ala Glu Lys Tyr Lys 35 40 45gat ttt aca aat tca gca gaa cat att gtt cca ggt cga ttt tat tta 552Asp Phe Thr Asn Ser Ala Glu His Ile Val Pro Gly Arg Phe Tyr Leu 50 55 60aaa gat tta aaa agt gga cta caa ttg aat aca tta gat cct aaa cga 600Lys Asp Leu Lys Ser Gly Leu Gln Leu Asn Thr Leu Asp Pro Lys Arg 65 70 75aaa ctt tat gta aca cga gaa ctt cat gga act gtt tta gtt gat tgt 648Lys Leu Tyr Val Thr Arg Glu Leu His Gly Thr Val Leu Val Asp Cys 80 85 90gtt gaa tta gtg gat gct gac atg gaa tgt aac tcg ggc atc ata cat 696Val Glu Leu Val Asp Ala Asp Met Glu Cys Asn Ser Gly Ile Ile His95 100 105 110aca att aga tat cct tta aca gat aca caa aaa cta gat aga tca tct 744Thr Ile Arg Tyr Pro Leu Thr Asp Thr Gln Lys Leu Asp Arg Ser Ser 115 120 125att ctc agc tta ctg gaa tcg aat tca gaa act caa tca ttt gca aaa 792Ile Leu Ser Leu Leu Glu Ser Asn Ser Glu Thr Gln Ser Phe Ala Lys 130 135 140gat tta tca gca gca ttg aaa aat aat ttg aaa aat att aac tct ggt 840Asp Leu Ser Ala Als Leu Lys Asn Asn Leu Lys Asn Ile Asn Ser Gly 145 150 155cat aga tac act gtg att gta cca agt caa aaa tcc tgg gaa tta tta 888His Arg Tyr Thr Val Ile Val Pro Ser Gln Lys Ser Trp Glu Leu Leu 160 165 170cga caa aag tat tca agt gaa caa cta caa aga att gca gct aat cat 936Arg Gln Lys Tyr Ser Ser Glu Gln Leu Gln Arg Ile Ala Ala Asn His175 180 185 190gtt atc ttg aat cag cat tgt tca gga cat tta att caa tca ctt gaa 984Val Ile Leu Asn Gln His Cys Ser Gly His Leu Ile Gln Ser Leu Glu 195 200 205aat tat aaa tca tta tta gga gaa aaa ctt gaa ttc att tgt gaa act 1032Asn Tyr Lys Ser Leu Leu Gly Glu Lys Leu Glu Phe Ile Cys Glu Thr 210 215 220gat tca act ggt aaa gaa acg aca tta tgt cgt act atg ttc aat gat 1080Asp Ser Thr Gly Lys Glu Arg His Tyr Val Arg Thr Met Phe Asn Asp 225 230 235aaa cat gaa tta aca ata aca gat tta acg tct tca aat gga gta gca 1128Lys His Glu Leu Thr Ile Thr Asp Leu Thr Ser Ser Asn Gly Val Ala 240 245 250cat att att gaa gaa cca tta att cca ttt tca gtt atg aca ttt aga 1176His Ile Ile Glu Glu Pro Leu Ile Pro Phe Ser Val Met Thr Phe Arg255 260 265 270gaa tta tta cag aac aag caa tat gct gaa aaa ttg aag tct act gaa 1224Glu Leu Leu Gln Asn Lys Gln Tyr Ala Glu Lys Leu Lys Ser Thr Glu 275 280 285ttt cta aaa ctt ctt gct gaa tgt gat ttg aaa gtg gaa cct gga gaa 1272Phe Leu Lys Leu Leu Ala Glu Cys Asp Leu Lys Val Glu Pro Gly Glu 290 295 300aaa tat gct atc gtt tta cca caa gac agt tca atg aag tgg tgg tca 1320Lys Tyr Ala Ile Val Leu Pro Gln Asp Ser Ser Met Lys Trp Trp Ser 305 310 315aca tat tca cca ttt caa gat gaa tat aaa cgt ttt caa gcg gat aaa 1368Thr Tyr Ser Pro Phe Gln Asp Glu Tyr Lys Arg Phe Gln Ala Asp Lys 320 325 330gaa tat cgt tgt cga gta gca cgt tac cat gtt gtg aaa agt gat aat 1416Glu Ser Arg Cys Arg Val Ala Arg Tyr His Val Val Lys Ser Asp Asn335 340 345 350cga ttg aat aat att ggc aat ttt gct agt cat aca cta ggt cat cgt 1464Arg Leu Asn Asn Ile Gly Asn Phe Ala Ser His Thr Leu Gly His Arg 355 360 365aca att aat cca aac gaa gct ctg tat gaa aca act tat ttt aag aag 1512Thr Ile Asn Pro Asn Glu Ala Leu Tyr Glu Thr Thr Tyr Phe Lys Lys 370 375 380act cca tat gga tca cag ctg aat ttc cat tat tct cca gtc aat aat 1560Thr Pro Tyr Gly Ser Gln Leu Asn Phe His Tyr Ser Pro Val Asn Asn 385 390 395ctt gaa tca tat gaa tta gat gat gtg aca atc tac gtt act cca agg 1608Leu Glu Ser Tyr Glu Leu Asp Asp Val Thr Ile Tyr Val Thr Pro Arg 400 405 410atc aat gtt cca cct gaa gta aat atg act gac ata ata aac aat cga 1656Ile Asn Val Pro Pro Glu Val Asn Met Thr Asp Ile Ile Asn Asn Arg415 420 425 430gct gat aca aca ata gcg aaa aat caa aca gaa aga gct aaa atg gat 1704Ala Asp Thr Thr Ile Ala Lys Asn Gln Thr Glu Arg Ala Lys Met Asp 435 440 445gaa aaa tat ttt aga aag aat gca cct aaa aat tta ttt ctt gtt aca 1752Glu Lys Tyr Phe Arg Lys Asn Ala Pro Lys Asn Leu Phe Leu Val Thr 450 455 460aca gat aat ggt tgg aag gat cca aga gct aca cag aca aca att aat 1800Thr Asp Asn Gly Trp Lys Asp Pro Arg Ala Thr Gln Thr Thr Ile Asn 465 470 475cca tat aaa cct gaa tta aat atg tat act gat aag aat tta gaa agt 1848Pro Tyr Lys Pro Glu Leu Asn Met Tyr Thr Asp Lys Asn Leu Glu Asn 480 485 490tac tta ctg ctt aac cat ata cca ttg tat tta tgg ggt gga gat att 1896Tyr Leu Leu Leu Asn His Ile Pro Leu Tyr Leu Trp Gly Gly Asp Ile495 500 505 510ggt tat ttc gaa aaa aat aca gta cat aaa ttt atg tct tct gct ggc 1944Gly Tyr Phe Glu Lys Asn Thr Val His Lys Phe Met Ser Ser Ala Gly 515 520 525ata gag tta atc ttt tgg atg gat aat aat gca gta atg cga att gga 2092Ile Glu Leu Ile Phe Trp Met Asp Asn Asn Ala Val Met Arg Ile Gly 530 535 540tat gag ggt cta cca cgt gat caa tgg cca aaa gtg att cat tgg aat 2040Tyr Glu Gly Leu Pro Arg Asp Gln Trp Pro Lys Val Ile His Trp Asn 545 550 555cta cat gca cgt gac ggt att atc tgg gta tta gat ggt cta ttg aaa 2088Leu His Ala Arg Asp Gly Ile Ile Trp Val Leu Asp Gly Leu Leu Lys 560 565 570tgt ccg gaa aaa ctt tgc cca tta ctt gct gaa gat gtt gat tat tat 2136Cys Pro Glu Lys Leu Cys Pro Leu Leu Ala Glu Asp Val Asp Tyr Tyr575 580 585 590gat gtt tat gta atg gca tgt caa aca tcc aat ttg ccg ggt gaa aaa 2184Asp Val Tyr Val Met Ala Cys Gln Thr Ser Asn Leu Pro Gly Glu Lys 595 600 605gat gct gtt gga caa ttt gaa agt aga cca ttg gat ata gct agt cgg 2232Asp Ala Val Gly Gln Phe Glu Ser Arg Pro Leu Asp Ile Ala Ser Arg 610 615 620cat cct aat ctc tgc act gtg gta ttg caa aca agt gaa aca acg gtt 2280His Pro Asn Leu Cys Thr Val Val Leu Gln Thr Ser Glu Thr Thr Val625 630 635aca ctt att gag tcc taa 2298Thr Leu Ile Glu Ser 640