[发明专利]日本血吸虫中国大陆株SjPP基因及其克隆、表达和应用

摘要

本发明涉及日本血吸虫中国大陆株SjPP基因的基因序列及其克隆表达方法和应用。本发明公开的日本血吸虫中国大陆株SjPP基因具有序列1所示的核苷酸序列和相应的序列2的氨基酸序列。本发明应用实验室繁殖的洞庭湖种群东方田鼠阳性血清和酶联葡球菌A蛋白免疫筛选日本血吸虫中国大陆株成虫cDNA文库，经过初筛和3轮复筛获得5个阳性克隆。应用5′RACE技术对其中一克隆的cDNA片段进行5′端延伸扩增，并利用blast进行基因序列的同源性分析，结果表明该基因为一血吸虫新基因。本发明还公开了SjPP在大肠杆菌中的表达方法。本发明SjPP基因可用于制备防治日本血吸虫病重组蛋白疫苗和DNA疫苗。

主权项

1、一种日本血吸虫中国大陆株SjPP基因，其特征在于该基因具有如下的核苷酸序列及相应的氨基酸序列：gggaatttgc ccttgca atg gac ttg gat cag tgg ata cag tct gtc aag 50 Met Asp Leu Asp Gln Trp Ile Gln Ser Val Lys 1 5 10act tgt aag tac ctt cca gaa aac gat ctt aga aaa ctg tgc aac tat 98Thr Cys Lys Tyr Leu Pro Glu Asn Asp Leu Arg Lys Leu Cys Asn Tyr 15 20 25gtt acg gag ttg ttg att gaa gag tgc aat gtg caa cca gtt tct ttt 146Val Thr Glu Leu Leu Ile Glu Glu Cys Asn Val Gln Pro Val Ser Phe 30 35 40cct gta acc ata tgt ggt gat att cat gga cag ttc tac gat ctt ttg 194Pro Val Thr Ile Cys Gly Asp Ile His Gly Gln Phe Tyr Asp Leu Leu 45 50 55cag tta ttt cgt cga gga gga aaa ctg ccg gac aaa aat tat ata ttt 242Gln Leu Phe Arg Arg Gly Gly Lys Leu Pro Asp Lys Asn Tyr Ile Phe60 65 70 75atg ggg gat ttt gtt gat cgg ggt tat tac agc ctc gaa acc ctc aca 290Met Gly Asp Phe Val Asp Arg Gly Tyr Tyr Ser Leu Glu Thr Leu Thr 80 85 90ttt ctt ctt gtt ctc aaa gct aag tat cct tca cga ata aca tta ttg 338Phe Leu Leu Val Leu Lys Ala Lys Tyr Pro Ser Arg Ile Thr Leu Leu 95 100 105aga gga aac cat gaa agt cgg caa gtt act cag tgt tat ggc ttc tat 386Arg Gly Asn His Glu Ser Arg Gln Val Thr Gln Cys Tyr Gly Phe Tyr 110 115 120gat gaa tgt atc aat aaa tat ggt aat gcg aat cca tgg cat tat tgt 434Asp Glu Cys Ile Asn Lys Tyr Gly Asn Ala Asn Pro Trp His Tyr Cys 125 130 135tgc aaa gtg ttc gat ctt ctc aca ata gcc gca tta gtt gga gaa aaa 482Cys Lys Val Phe Asp Leu Leu Thr Ile Ala Ala Leu Val Gly Glu Lys140 145 150 155gtt ctc tgc gtt cat gga ggc cta tca cct caa att aaa aca ctc gat 530Val Leu Cys Val His Gly Gly Leu Ser Pro Gln Ile Lys Thr Leu Asp 160 165 170caa atc cgg aca atc gac aga cat atg gaa ata ccc aat gaa ggt cca 578Gln Ile Arg Thr Ile Asp Arg His Met Glu Ile Pro Asn Glu Gly Pro 175 180 185ttt tgt gac att tta tgg tcg gat cct gat gac gtg gct cat tgg gct 626Phe Cys Asp Ile Leu Trp Ser Asp Pro Asp Asp Val Ala His Trp Ala 190 195 200gtt agt ccg cgt gga gca gga tat tta ttt ggg cca aaa gtt act aat 674Val Ser Pro Arg Gly Ala Gly Tyr Leu Phe Gly Pro Lys Val Thr Asn 205 210 215gaa ttc gtt gag tta aat ggt cta gaa tgt att tgt aga gct cat caa 722Glu Phe Val Glu Leu Asn Gly Leu Glu Cys Ile Cys Arg Ala His Gln220 225 230 235ctc gtg cac gaa ggc tac aag tgc atg ttc gat gga caa cta atc act 770Leu Val His Glu Gly Tyr Lys Cys Met Phe Asp Gly Gln Leu Ile Thr 240 245 250att tgg tca gct cca aat tat tgt tat cgt tgt gga aat gtt gca gcg 818Ile Trp Ser Ala Pro Asn Tyr Cys Tyr Arg Cys Gly Asn Val Ala Ala 255 260 265att ttg gaa ttc caa agt cca aca caa ttc gat cct aaa ctt ttt gaa 866Ile Leu Glu Phe Gln Ser Pro Thr Gln Phe Asp Pro Lys Leu Phe Glu 270 275 280gct gtg cca gat gaa gaa cga gtc aaa cct cct cta agg att act ccg 914Ala Val Pro Asp Glu Glu Arg Val Lys Pro Pro Leu Arg Ile Thr Pro 285 290 295tac ttc ttg taacatattc ttcttaaact tgatcatttt cattctaagt 963Tyr Phe Leu300acaagttttc ttgttaactc cgattaacac tgcgatatcc aacagcaata ggaacattta 1023tttgcattct aaatccagtt gtacagattt gagtccaatt atttggttct catacctaaa 1083tgtatgtatt ctttttgcaa tttttattta atcgaaatgt atcaccgaca ctgcaatgat 1143tttgtctttc tgacggatcc aacagactgg taattgatga atatacacag catgcggaca 1203attggaagag taaaattttc gcttaacaca ccagacaagt tttctcggac tagtgtgacg 1263ataatacata catcttatcc caaaaaaaaa aaaaaaaaaa aaaaaaaagg gcggcg 1319