[发明专利]日本血吸虫HGPRT基因的克隆、表达和DNA疫苗

摘要

本发明涉及生物基因领域，具体涉及日本血吸虫(中国大陆株)HGPRT基因的基因序列、原核表达质粒pGEX－4T－1/SjHGPRT的构建及鉴定、真核表达质粒pcDNA3/HGPRT的构建及重组质粒免疫小鼠的保护性效果观察。其发明基于我们对HGPRT基因可以作为抗日本血吸虫病天然分子基因疫苗研究靶标的发现，根据SjHGPRT的cDNA序列的阅读框(ORF)设计引物，并从日本血吸虫成虫cDNA文库中调出该基因片段的完整阅读框，然后亚克隆入真核表达载体pcDNA3中，构建成重组质粒pcDNA3/SjHGPRT，制备DNA疫苗免疫昆明系小鼠，获得30.4％的减虫率、44.8％的减卵率。

主权项

1、一种编码日本血吸虫(中国大陆株)HGPRT基因的核苷酸序列，其特征在于它具有如下核苷酸序列： cgaacatcttcacgac 16atg tct ggt gtt atg aaa aga act ggt tgt att gtg ata gaa gat 61Met Ser Gly Val Met Lys Arg Thr Gly Cys Ile Val Ile Glu Asp 5 10 15gac ttc aaa ggt ttt cct att gag cat ttt tgc acg tct cct agg 106Asp Phe Lys Gly Phe Pro Ile Glu His Phe Cys Thr Ser Pro Arg 20 25 30tat gag ggt tgt ttg gac tac gtt ctc gta tca aac ggt atg atc 151Tyr Glu Gly Cys Leu Asp Tyr Val Leu Val Ser Asn Gly Met Ile 35 40 45aag gat aga ctt gaa aaa atg tca atg gat att gtt gac tct tat 196Lys Asp Arg Leu Glu Lys Met Ser Met Asp Ile Val Asp Ser Tyr 50 55 60gaa gct tct aat gca aca tct atc aca ctt atg tgt gtt ctg aaa 241Glu Ala Ser Asn Ala Thr Ser Ile Thr Leu Met Cys Val Leu Lys 65 70 75ggt gga ttc aaa ttt ctt gct gat ctt gtc gat ggt ctt gag cgt 286Gly Gly Phe Lys Phe Leu Ala Asp Leu Val Asp Gly Leu Glu Arg 80 85 90act gta cgt gct cga ggt atc gtc ctc cct atg tca gtt gaa ttc 331Thr Val Arg Ala Arg Gly Ile Val Leu Phe Met Ser Val Glu Phe 95 100 105gtt cgc gtt aaa agc tat gtt aat gat gtc agc tat cat gag cct 376Val Arg Val Lys Ser Tyr Val Asn Asp Val Ser Tyr His Glu Phe 110 115 120ttg tta act ggt ttg ggg gac cct tca gac tac aga gat aag gat 421Lys Leu Thr Gly Leu Gly Asp Phe Ser Asp Tyr Arg Asp Lys Asp 125 130 135att ctt gtt gta gaa gat ata atc gat acg ggc aga aca ata acc 466Ile Leu Val Val Glu Asp Ile Ile Asp Thr Gly Arg Thr Ile Thr 140 145 150aaa ttg ata aat cac ttg gaa agc ttg tcc acg aaa agt gtc aag 511Lys Leu Ile Asn His Leu Glu Ser Leu Ser Thr Lys Ser Val Lys 155 160 165gta gca agc ctt ctc gtc aaa cgt aca atg cct ccc agt cag tat 556Val Ala Ser Leu Lys Val Lys Arg Thr Met Ppo Pro Ser Glu Tyr 170 175 180cat cca gat ttt gtt ggc ttt gaa gtt ccg aat cgg ttt gtt gtt 601His Pro Asp Phe Val Gly Phe Glu Val Pro Asn Arg Phe Val Val 185 190 195ggt tat gct tta gac tac aat gaa cat ttc cgt gac cta cac cat 646Gly Tyr Ala Leu Asp Tyr Asn Glu His Phe Arg Asp Leu His His 200 205 210att tgt gtg att aat gaa att ggt caa caa aaa ttc tct gta cct 691Ile Cys Val Ile Asn Glu Ile Gly Gln Gln Lys Phe Ser Val Pro 215 220 225tgt gtg ccg aaa tca gtt 709Cys Val Pro Lys Ser Val 231taatgttcaa aagatcggat acacatataa taatgttcaa aagatcggat 759acacatataa acaaggatta ttcatagtgc acaatctcta aatgaacaca 809atgtaatgat taacttattt tgggaaggaa aaacatttac ttcattcaca 859ctgttcatgc aaccatatgt ttccgtgttt atgtctgtat cttacagttt 909ctaactacgt cgtgtatttt taatttttgt tttcatgata tgctctgtat 959tatgaacttc taatttttcc acattaatta ttttgttcac tttttttgat 1009atcgttcttt attctctact tgtcatattt tctgtttaga aggtcataca 1059tacacgtatt tacctattaa tcgacattgt ttttctgatt catacgtctt 1109taagctattt tctttttaaa agtctggttg tcgccacgtt gttcatattg 1159atgttgtgat ggttggtact atatagtcat tttgtttgct ccttgttatt 1209cactgaagta gtaaagtatt cttgtgatta ttattaaatg taaacttggt 1259aaaataagat taaatggaat tggatatgaa aaaaaaaaaa a 1300。