日本血吸虫普遍性结合酶E基因的获得和分析

目的从日本血吸虫(Schistosoma japonicum)成虫cDNA文库中获得并分析日本血吸虫新基因,为日本血吸虫病的防治提供候选疫苗和药物靶点.方法构建日本血吸虫成虫cDNA文库,随机挑取重组阳性克隆进行测序,对部分序列进行步移法测序获取全长cDNA,登录GenBank,利用BLAST程序对同源性高的基因进行核苷酸和氨基酸水平的同源性比较;并利用pcgene软件对其蛋白质结构进行初步分析.结果获得了1个日本血吸虫新基因普遍性结合酶E基因(ubiqui-tin-conjugating enzyme E,AY251608),长854bp,编码149个氨基酸,与人类普遍性结合酶E具有55%的同源性,编码蛋白的理论分子量为7.149 8 kDa,等电点为8.01;抗原表位可能位于氨基酸序列373～396处.结论步移法测序和生物信息学技术相结合有利于发现日本血吸虫新基因.

ACQUIRING AND ANALYZING OF UBIQUITIN-CONJUGATING ENZYME E GENE OF SCHISTOSOMA JAPONICUM

Objective To acquire and analyze Schistosoma japonicum adult stage new genes from an adult Schistosoma japonicum cDNA library and to search new vaccine candidates and drug targets for schistosomasis control. Methods A cDNA library was constructed from Shistosoma japonicum (Chinese strain) adult stage. Clones were selected randomly from the cDNA library and sequenced. Novel genes were acquired after analysis in GenBank databases by BLAST programs and other programs,and the analysis of protein was carried out with pcgene software. Results Ubiquitin-conjugating enzyme E gene,854 base pair and coding for 149 amino acids,was acquired and was submitted to GenBank and to get an accession number that was AY251608. This gene was homologous with human ubiquitin-conjugating enzyme E gene with 55% identity,The academic molecular weight was 7.149 8 kDa,pI 8.01,and the antigen determinant probably from 373 to 396 on the cDNA sequence. Conclusion It is a feasible method to discover and acquire novel genes of Schistosoma japonicum by combination of the Primer-walking sequencing and Bioinformatics technique.