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

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(英文)

作　　者：程国锋[1] 林矫矫[1] 孙军[1] 李浩[1] 朱传刚[1] 周元聪[2] 蔡幼民[1] 

机构地区：[1]中国农业科学院上海家畜寄生虫病研究所农业部动物寄生虫学重点实验室,上海200232 [2]中国科学院上海生命科学研究院上海生化细胞所,上海200031

出　　处：《Zoological Research》2005年第5期513-517,共5页动物学研究（英文）

基　　金：国家高技术研究发展项目"863"计划(2001AA215151);上海市重大科技攻关项目(03DZ19231)

摘　　要：The expressed sequence tag of eukaryotic translation initiation factor 5 （eIF5） from the Schistosoma japonicum adult worm cDNA library through subtractive hybridization between male and female worms was analyzed by the bioinformatics method. The overlapping sequences were assembled into one that includes the complete open reading frame （GenBank accession number： AY686501）. The full-length cDNA of SjeIF5 was cloned into a pET-28c^（＋） vector, which generated a prokaryotic expression plasmid, and a fusion protein of 18 kDa was induced in Escherichia coll. The recombinant 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.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。

关 键 词：Schistosoma japonicum eIF5 EXPRESSION VACCINE 

分 类 号：R392[医药卫生—免疫学]