【摘要】 利用快速cDNA末端扩增(RACE)技术结合“电子cDNA文库”筛选法,对日本血吸虫成虫期特异表达基因EST片段的全长序列进行了扩增,获得了一含完整开放性阅读框(ORF)的基因序列(GenBank中的登录号为AY847290)。经对该基因序列进行分析,表明其为日本血吸虫的一新基因,命名为Sj314C10。将该基因的蛋白编码区克隆到真核表达载体pcDNA3 中,把构建好的重组质粒转化到大肠埃希氏菌DH5α中培养;筛选阳性克隆进行序列鉴定,经分析表明与预期的结果一致。成功地构建了该基因的DNA疫苗,为该基因作为疫苗候选分子的研究奠定了基础。更多还原

【Abstract】 The rapid amplification of cDNA ends(RACE)technique and the electronic cDNA library select mathod was applied to amplify full length of the expressing sequence tag(EST) ,which is a specfic-stage in Schistosoma japonicum adult worm, and a cDNA fragment with an ORF was identified(GenBank accession number is AY847290). The bioinformatics sequencing revealed that the gene was a novel gene of S.japonicum, and it was named as Sj314C10(S.japonicum 314C10 cloning).The codons protein region of the gene was cloned into pcDNA3-vector and recombinant plasmid was transformed into E.coli DH5α. Positive clone of recombinant plasmid bacterial E.coli DH5α was selected to identify. The result (agreed) with what was anticipated.Thus a recombinant expression plasmid was constructed successfully. It was made foundation for further studying the gene’s function.更多还原