鹅细小病毒SS/10株VP3蛋白的原核表达及鉴定  

Prokaryotic Expression and Identification of VP3 Gene of Goose Parvovirus SS/10 Strain

作　　者：胡自立[1] 刘海玲[1] 张光明[1] 张凯[1] 高瑞娟[1] 罗开健[1] 

机构地区：[1]华南农业大学兽医学院农业部兽用疫苗创制重点实验室,广东广州510642

出　　处：《动物医学进展》2012年第5期35-38,共4页Progress In Veterinary Medicine

基　　金：农业科技成果转化资金项目(2011GB2E000005)

摘　　要：根据GenBank收录的鹅细小病毒(GPV)基因序列,设计并合成了一对VP3基因扩增引物。采用PCR技术对SS/10株的VP3基因进行扩增,将目的基因和原核表达载体PET-32a分别经HindⅢ和BamHⅠ双酶切后进行连接,获得重组质粒PET-32a-VP3,并将其转化表达菌BL21(DE3)pLysS。经IPTG诱导,SDS-PAGE分析,结果获得了72ku左右的融合蛋白,大部分以包涵体的形式存在于菌体中。Wesern blot结果表明,该蛋白能与GPV阳性血清发生特异性反应,具有良好的反应原性。One pair of primers was designed to amplify VP3 gene according to the sequences of goose parvovirus（GPV）published in GenBank.The PCR product of SS/10 was cloned into the expression vector PET-32a after double enzyme digestion by HindⅢ and BamHⅠ.The recombinant prokaryotic expression plasmid was constructed,and named PET-32a-VP3.The recombinant plasmid was transformed into BL21（DE3） pLysS.The transformed bacteria were induced with IPTG and the expressed protein was analyzed by SDS-PAGE.The fusion protein of 72 ku was expressed.The protein could react with GPV positive serum specifically in a Western blot test,indicating that the expressed protein had good reactogenicity.

关 键 词：鹅细小病毒 VP3基因 原核表达 

分 类 号：S852.659.2[农业科学—基础兽医学]