细粒棘球绦虫G1型Eg95基因的原核表达及蛋白鉴定

Prokaryotic Expression and Protein Identification of Eg95 Gene from G1 Genotype of Echinococcus granulosus

根据GenBank发表的细粒棘球绦虫G1型Eg95序列合成Eg95抗原基因(HM345604.1),设计并合成一对引物,采用PCR技术扩增Eg95抗原基因,亚克隆到pET-28a原核表达载体,并在大肠埃希菌BL21(DE3)中表达重组蛋白。经IPTG诱导,SDS-PAGE分析,Eg95抗原蛋白可以在大肠埃希菌中高效表达,表达重组蛋白的分子质量约为16.5ku。Western blot结果表明,该蛋白可与阳性血清发生特异反应,具有很好的反应原性。

Eg95 antigen gene was synthetized according to the sequences published in the GenBank of Eg95 gene from G1 genotype of Echinococcus granulosus, a pair of specific primers were designed and synthe- sized. The fragment was cloned into pET-28a vector. The recombinant plasmid was then transformed into E. coli BL21 （DE3）. The recombinant protein was analyzed by SDS-PAGE, the result showed that recombinant protein was highly expressed in E. coli BL21 （DE3） and the molecular weight was 16.5 ku. It could be recognized by positive serum specifically, and showed good activity in immune response.