细粒棘球绦虫六钩蚴EG95抗原基因的克隆及原核表达  

Cloning and prokaryotic expression of EG95 gene of Echinococcus granulosus oncospheres

作　　者：韩秀敏[1] 贾万忠[1] 史大中[2] 

机构地区：[1]中国农业科学院兰州兽医研究所 [2]兰州医学院,甘肃兰州730000

出　　处：《中国兽医科技》2005年第5期381-385,共5页Chinese Journal of Veterinary Science and Technology

基　　金：瑞典国际基金项目(B/2857 1)

摘　　要：利用RT PCR从激活的细粒棘球绦虫六钩蚴中扩增了EG95 cDNA,并亚克隆至原核表达载体pGEX 4T 1上,转化至大肠埃希氏菌BL21,用IPTG诱导重组菌株表达了融合蛋白质。结果表明,从六钩蚴中克隆到了EG95 cDNA,序列长度为481 bp,包括471 bp的开放阅读框,与新西兰株EG95抗原基因序列完全一致;SDS PAGE电泳结果显示,重组表达质粒产生了约42 ku的目的带,其中EG95蛋白质的分子质量约为15.2 ku,与预期结果一致;Western blotting试验检测表明,融合表达产物可与囊性包虫病人血清发生反应。试验结果提示,EG95 基因高度保守,所表达的EG95蛋白作为抗原疫苗具有广泛的适用性。The EG95 cDNA fragments were amplified from the hatched and activated oncospheres (Echinococcus) granulosus by RT-PCR, and then subcloned into pGEX-4T-1 vector and transformed into (E.coli) BL21. The recombinant transformants were induced with IPTG. The results showed that the EG95 cDNA with a 471bp of open reading frame, was 481bp in length and the sequence of which was the same as that from the New Zealand strain. The SDS-PAGE electrophoresis indicated that the molecular weight of the expressed fusion protein in recombinant E.coli was approximately 42ku, and the molecular weight of the EG95 in the fusion protein was about 15.2ku, which were the same as the expected results. The Western-blotting test showed that the fusion protein could be recognized by the positive sera from echinococcosis patients. It was concluded that the EG95 cDNA is conservative and can be used as a recombinant vaccine.

关 键 词：细粒棘球绦虫 EG95基因 克隆 原核表达 

分 类 号：S852.734[农业科学—基础兽医学] Q786[农业科学—兽医学]