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

作　　者：徐丹[1,2] 曹利利 魏峰 杨美英[1] 张莹光[1,2] 商立民[2] 刘全[2] 

机构地区：[1]吉林农业大学生命科学学院,吉林长春130118 [2]军事医学科学院军事兽医研究所吉林省人兽共患病预防与控制重点实验室,吉林长春130122 [3]吉林省畜牧兽医科学研究院,吉林长春130062

出　　处：《动物医学进展》2013年第4期46-49,共4页Progress In Veterinary Medicine

基　　金：国家重点基础研究发展计划(2012CB722501);军事医学科学院创新基金(2012CXJJ019)

摘　　要：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.

关 键 词：细粒棘球绦虫 EG95 原核表达 Western BLOT分析 

分 类 号：S855.99[农业科学—临床兽医学]