鹅细小病毒VP3基因的克隆及表达  

Cloning and Expression of VP3 Gene of Goose Parvovirus

作　　者：孙敏华[1] 董嘉文[1] 李林林[1] 袁建丰[1] 邝瑞欢[1] 胡奇林[1] 

机构地区：[1]广东省农业科学院动物卫生研究所,广东省公共卫生公共实验室,广东广州510640

出　　处：《广东畜牧兽医科技》2013年第5期25-28,共4页Guangdong Journal of Animal and Veterinary Science

基　　金：广东省科技计划项目(2012A020100001);广东省兽医公共卫生公共实验室开放基金项目(GSKJ090201)

摘　　要：根据GenBank中的鹅细小病毒(GPV)全基因序列,设计并合成了1对特异性引物,利用PCR对GPV-Fos han-2009株的VP3基因进行了扩增。经酶切、连接、转化后,获得了pET32a-VP3重组表达载体。经BL21(DE3)原核表达显示,VP3蛋白为包涵体形式,37℃下该蛋白最佳I PTG诱导浓度为1.2 mM,最佳诱导时间为5 h。SDS-PAGE分析显示所表达的VP3融合蛋白的相对分子质量约80 kD。West ern Bl ot证实该蛋白能与番鸭抗GPV阳性血清反应,这为GPV血清学诊断方法的建立提供了物质基础。A pair of primers based on the complete genome sequences of Goose Parvovirus were designed to amplify the VP3 gene of GPV-Foshan-2009 by PCR method. After enzyme digestion, ligation and tranduction, recombinant pET32a-VP3 vecter was constructed. The fusion protein in the form of inclusion bodies was highly expressed in E.coli BL21 （DE3）induced by 37℃ ,1.2 mM IPTG for 5 hours. The molecular weight of recombinant fusion protein is 80 KDa. The recombinant protein exhibited positive reactivity with the GPV antibodies in western-blot test, as would be helpful for the serological diagnosis research.

关 键 词：鹅细小病毒 VP3基因 克隆 表达 

分 类 号：S852.659.5[农业科学—基础兽医学]