[发明专利]日本血吸虫SJCXWL02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)SjCXWL02基因的基因序列、真核表达质粒pcDNA3/SjCXWL02的构建及重组质粒免疫小鼠的保护性效果观察。采用日本血吸虫致弱尾蚴血清筛选尾蚴cDNA文库，获得8个阳性克隆后，PCR扩增出其中4个含完整ORF的基因。同源性序列分析表明其中一个为新基因(SjCXWL02)，该基因全长1879bp，开放阅读框含984bp，编码328个氨基酸。将SjCXWL02亚克隆入真核表达载体pcDNA3中，构建成重组质粒pcDNA3/SjCXWL02，制备DNA疫苗免疫昆明系小鼠，获得24.6％的减虫率、32.0％的减卵率。

主权项

1、一种编码日本血吸虫SJCXWL02基因的核苷酸序列，其特征在于它具有如下核苷酸序列： 1 ggggaatatagtttcgattttctcgtgggacgatggcaggggatgataaa Met Ala Gly Asp Asp Lys 51 atgaatattgatagtatcattgccagattactggaggtccgacgcattcgg Met Asn Ile Asp Ser Ile Ile Asp Arg Leu Leu Glu Val Arg Arg Ile Arg102 cccggcaaaaatgttcaactgaacgaggcagaaattcgcggattgtgtctt Pro Gly Lys Asn Val Gln Leu Asn Glu Asn Glu Ile Arg Gly Leu Cys Ile153 aaatctcgcgagattttcttaagtcaacccattttactggaattagaggct Lys Ser Arg Glu Ile Phe Leu Ser Gln Phe Ile Leu Leu Glu Leu Glu Ala204 cccctcaaaatttgtggtgacattcacggccaatattatgatttacttcga Pro Leu Lys Ile Cys Gly Asp Ile His Gly Gln Tyr Tyr Asp Leu Leu Arg255 ctttttgaatatggagcgtttcctcctgaagcgaattatctgtttttgggt Leu Phe Glu Tyr Gly Ala Phe Pro Pro Glu Ala Asn Tyr Leu Phe Ile Gly306 gactatgtggatcgagggaagcagtcactggaaacaatatgcttattatta Asp Tyr Val Asp Arg Gly Lys Gln Ser Leu Glu Thr Ile Cys Leu Ile Ile357 gcttataaaatcaagtatccggaaaatttctttctgcttaggggtaatcat Ala Tyr Lys Ile Lys Tyr Pro Glu Asn Phe Phe Leu Leu Arg Gly Asn His408 gaatgtgcctccatcaaccgtatatatggattttatgatgaatgtaaacgt Glu Cys Ala Ser Ile Asn Arg Ile Tyr Gly Phe Tyr Asp Glu Cys Lys Arg459 cgatataatataaaattgtggaagaccttcacagactgcttcaactgtttg Arg Tyr Asn Ile Lys Leu Trp Lys Thr Phe Thr Asp Cys Phe Asn Cys Leu510 ccgattgtagcgattatagacgagaaaatattttgctgtcatggtggtctc Pro Ile Val Ala Ile Ile Asp Glu Lys Ile Phe Cys Cys His Gly Gly Leu561 tcaccagatctttcaacaatggaacaaataagacgaatcatgcgtcctaca Ser Pro Asp Leu Ser Thr Met Glu Gln Ile Arg Arg Ile Met Arg Pro Thr612 gatgttcctgagcaaggtttactatgtgatttactttggtcagatccagac Asp Val Pro Glu Gln Gly Leu Leu Cys Asp Leu Leu Trp Ser Asp Pro Asp 663 aaagatgtatctggttggggtgaaaatgatcgtggtgtcagttatacattt Lys Asp Val Ser Gly Trp Gly Glu Asn Asp Arg Gly Val Ser Tyr Thr Phe 714 ggtccagatattgtagctagattcttacataaacatgatctggatttaata Gly Pro Asp Ile Val Ala Arg Phe Leu His Lys His Asp Leu Asp Leu Ile 765 tgtcgtgctcatcaggttgttgaagacggttatgaa ttcttcgctaaacgt Cys Arg Ala His Gln Val Val Glu Asp Gly Tyr Glu Phe Phe Ala Lys Arg 816 cagttagttacactgttttccgctccaaattattgtggtgaattcgataat Gln Leu Val Thr Leu Phe Ser Ala Pro Asn Tyr Cys Gly Glu Phe Asp Asn 867 gcaggagctatgatgtctgtagatgacacattattatgttcatttcaaatt Ala Gly Ala Met Met Ser Val Asp Asp Thr Leu Leu Cys Ser Phe Gln Ile 918 ttacgcccagctgaaaagaaaaagtattacgctggtgtcccaactggcagt Leu Arg Pro Ala Glu Lys Lys Lys Tyr Tyr Ala Gly Val Pro Thr Gly Ser 979 cgaccaattactccacctcgtggagctaaagctaaggggaaattataa Arg Pro Ile Thr Pro Pro Arg Gly Ala Lys Ala Lys Gly Lys Leu *1010 actatctatc tcatcgcgta tctctccaat ttatttgctt1050 ttgcttttat gacaataaaa tcgaggcgtc aagtggacga1090 acaaagttat ttgagaatat caccccctgt tcaaactgtc1130 attaaaatca tttttgcgtc ataaagtgta gatttccctt1170 acatgtgtct ttttcggaat ggcaaatgtt tttttggtcc1210 tgcacatcaa gattggataa tcagtgtgaa atcctgtttt1250 tagtgtttat acgttgttaa acagccttca tgtttccctt1290 tctatactta tttacaagcc tcatccttat cgactggctt1330 gttctctttc ctttttgcat gttttgtaag caagcataca1370 caaaagattg gttgttgatt gtgtcctatt tagtagtacc1410 atttttatta ccactggtat aattgtacat ttcttcttcg1450 tatgttttat tcttattgaa atattcaaat taaagaatga1490 gttgttttta tcagtttcgc tcaacattat tcatttgatt1530 tgatcattca gtcagtccag tgtgtaaaac acataattta1570 acagtagaaa ctgtttacat gtagtagttt ttataaatca1610 caagcatctt tgtttcttct gtcagtattt atttgtttgt1650 gtgtatgtga ttatagtgat gatgaatgtg tgcatgctgt1690 tttttcttgt cttcgaattc gattgttcac aatgataact1730 gaagtttttc acattttctg ttttcccaat ttacttttat1770 tgtacagtca gctactttct gttataaaaa aacctatcct1810 tagtatgtta aattgagtga aatataattt cacctttcga1850 aaaaaaaaaa aaaaaaaaaa aaaaaaaaa。