鹿流行性出血病病毒VP7基因的克隆及原核表达  

Cloning and Prokaryotic Expression of VP7 Gene of Epizootic Hemorrhagic Disease Virus of Deer in E.coli

作　　者：杨俊兴[1,2] 花群义[1] 陈焕春[2] 陈兵[1] 吕建强[1] 曹琛福[1] 阮周曦[1] 张彩红[1] 

机构地区：[1]深圳出入境检验检疫局动植物检验检疫技术中心,广东深圳518001 [2]华中农业大学动物医学院,湖北武汉430070

出　　处：《动物医学进展》2009年第8期5-8,共4页Progress In Veterinary Medicine

基　　金：国家863计划项目(2006AA10Z445);中国博士后科学基金项目(20080430855)

摘　　要：根据已经扩增的鹿流行性出血病病毒(EHDV)VP7基因序列,设计一对扩增VP7基因特异性引物,用RT-PCR从血清I型EHDV(EHDV-1)总RNA中扩增VP7基因,并将其克隆到原核表达载体pBAD/Thio中,构建了重组原核表达质粒pBAD/Thio-EHDV VP7。将表达载体转入大肠埃希菌(E.coliLMG194),用L-阿拉伯糖诱导表达。SDS-PAGE和Western blot试验结果表明,EHDV VP7基因在LMG194中得到了表达,其表达产物可以与EHDV阳性血清特异性反应,说明该融和蛋白具有免疫学活性。According to the published sequence in GenBank of VP7 gene of epizootic hemorrhagic disease of deer virus （EHDV）, a pair of specific primers were designed and synthesized. The VP7 gene was amplified by RT-PCR from the total RNA extracted from EHDV. The PCR product was cloned into pBAD/Thio liner plasmid forming the recombinant expression vector pBAD/Thio-EHDV VP7. The pBAD-EHDV-VP7 was transformed into E. coli LMG194 and induced by L-arabinose to express recombinant protein. SDS- PAGE and Western blot indicated that recombinant VP7 protein was expressed successfully with molecular weight of 54.5 ku and could specifically bind to positive sera anti EHDV.

关 键 词：鹿流行性出血病病毒(EHDV) VP7基因 基因克隆 表达 

分 类 号：S859.659.4[农业科学—临床兽医学]