日本血吸虫真核翻译起始因子5基因的cDNA克隆和免疫保护功能分析

利用抑制削减杂交法筛选日本血吸虫雌雄虫差异基因,从中获得真核翻译起始因子5基因(eIF5)的 表达序列标。对该序列进行生物信息学分析:对其5’端进行电子延伸,获得含完整开放阅读框的cDNA序列 (AY686501)。将其亚克隆到表达载体pET-28c,进行重组表达。免疫保护效果实验表明,重组表达蛋白具有显著 抑制血吸虫卵在寄主肝脏中的沉积效果。

Cloning and Functional Analysis of the Full Length cDNA Sequence of Eukaryotic Translation Initiation Factor 5 in Schistosoma japonicum

The expressed sequence tag of eukaryotic translation initiation factor 5 (elF5) from the Schistosoma japonicum adult worm cDNA library through subtractive hybridization between male and female worms was analyzed by the bioin-formatics method. The overlapping sequences were assembled into one that includes the complete open reading frame (GenBank accession number: AY686501). The full-length cDNA of SjelF5 was cloned into a pET-28c( ) vector, which generated a prokaryotic expression plasmid, and a fusion protein of 18 kDa was induced in Escherichia coli. The recombi-nant expression of eIF5 protein of Schistosoma japonicum was purified. The immunoprotection test against schistosomiasis demonstrated that the recombinant protein worked to a certain extent, especially in the reduction of eggs in the liver of the host.