猪圆环病毒II型衣壳蛋白片段与T4噬菌体SOC蛋白的融合表达  

Fused Expression of PCV2 Cap Gene and T4 Phage SOC Gene

作　　者：王宪文[1] 毕英佐[1] 王新卫[1] 焦玉萍[1] 谢青梅[1] 曹永长[1] 于康震[1] 

机构地区：[1]华南农业大学动物科学院

出　　处：《中国农学通报》2008年第5期58-61,共4页Chinese Agricultural Science Bulletin

基　　金：广东省科技重点攻关项目A20403002

摘　　要：将PCV2衣壳蛋白片段与T4噬菌体SOC蛋白在大肠杆菌中融合表达。利用PCR技术扩增猪圆环病毒Cap基因。将Cap基因克隆至T4噬菌体表达质粒pSOC的SOC基因(T4噬菌体表面非结构蛋白基因)下游,构建成T4噬菌体SOC位点表达Cap的表达载体pSOC-Cap。将其转化至大肠杆菌BL21(DE3),经IPTG诱导后表达的目的蛋白SOC-Cap进行SDS-PAGE与Western-blot方法检测。表达的SOC-Cap蛋白相对分子质量约28kD,表达量占菌体总蛋白量的22.31%,并具有与PCV2特异性抗体反应的活性。PCV2衣壳蛋白片段与T4噬菌体SOC蛋白的融合表达后,表达量较高,并具有免疫活性,有望用于猪圆环病毒II型的诊断和预防。The PCV2 Cap gene and T4 Phage SOC gene was expressed fusedly in E.col. Cap gene of PCV2 was amplified by PCR and cloned into the expressing plasmid pSOC, where Cap gene was fused to 3＇end of T4 phage small outer capsid gene encoding SOC protein. T4 phage expression vector named pSOC-Cap was constructed and used to transform E.coli BL21（DE3）. The combinant E.coli BL21（DE3） was induced by IPTG. The expressed fusion protein SOC-Cap was detected by SDS-PAGE with expected molecular size of 28kD and the expression product accounted for 22.31% of the total bacterial protein. The immunological test applying Western-blot indicated that SOC-Cap fusion protein could react to PCV2 specifically. The SOC-Cap fusion protein expressed successfully with high production and immunocompetence could be used to diagnose and prevent PCV2.

关 键 词：猪圆环病毒II型 衣壳蛋白 T4噬菌体 融合表达 

分 类 号：S852.659[农业科学—基础兽医学]