表达猪圆环病毒2型Cap蛋白的重组杆状病毒的构建及鉴定  

Construction and Identification of a Recombinant Baculovirus Expressing Cap Protein of Porcine Circovirus Type 2

作　　者：张永武[1] 连海[1] 张锦霞[1] 张静远[1] 刘晔[1] 张守峰[1] 扈荣良[1] 

机构地区：[1]军事医学科学院军事兽医研究所,吉林长春130000

出　　处：《动物医学进展》2016年第3期14-18,共5页Progress In Veterinary Medicine

基　　金：国家自然科学基金项目(31372456;31472214)

摘　　要：利用Bac-to-Bac杆状病毒表达系统成功表达了猪圆环病毒2型Cap蛋白,并对该蛋白的生物活性进行鉴定。PCR获得Cap基因并克隆入pFastBacTMⅠ载体质粒,重组质粒pFastBacⅠ-Cap转化DH10Bac感受态细胞,通过蓝白斑筛选获得重组杆粒rBacmid-Cap。以脂质体法将重组杆粒转染对数生长期的Sf9细胞,获得重组杆状病毒。电镜下可见典型的杆状病毒,SDS-PAGE显示,重组病毒表达的Cap蛋白约28ku,间接免疫荧光试验证明,Cap蛋白良好表达并具有免疫反应性,为猪圆环病毒亚单位疫苗的研究奠定了基础。The study utilized Bac-to-Bac（R） baculovirus expression system to generate Cap protein of porcine circovirus type 2and identified the biological activity of Cap protein.Cap gene was cloned into pFastBacTMⅠ.The recombinant plasmid pFastBacⅠ-Cap was transformed into DH10 Bac,and the recombinant rBacmid-Cap was obtained by blue-white screening.Sf9 cells were transfected with rBacmid-Cap by liposome method and recombinant baculovirus rAcMNPV-Cap was constructed.Baculovirus was observed by electron microscope.Cap protein was about 28 ku which was confirmed by SDS-PAGE.The indirect immunofluorescence assay showed that Cap protein had good immunogenicity.This study laid a foundation for the development of the subunit vaccine of porcine circovirus type 2.

关 键 词：猪圆环病毒2型 CAP蛋白 杆状病毒表达系统 

分 类 号：S852.659.2[农业科学—基础兽医学]