作　　者： ; ; ; ;

机构地区： 华南农业大学兽医学院

出　　处： 《中国畜牧兽医》 2013年第7期6-10,共5页

摘　　要： 通过PCR方法扩增猪圆环病毒2型(porcine circovirus 2,PCV2)核定位序列部分缺失的ORF2基因(pdCap),将其克隆至Bac-to-Bac杆状病毒表达系统的pBACsurf-1质粒中,得到重组质粒pBACsurf-1-pdCap。再将含有pdCap和gp64的融合基因克隆到杆状病毒并转移到质粒pFastBacTM Dual中,得到重组质粒pFastBac-pdCap。然后将该重组质粒转化至DH10Bac感受态细胞中,经蓝白斑筛选和PCR鉴定,得到重组穿梭质粒Bacmid-pdCap。在脂质体介导下转染Sf9昆虫细胞,获得重组杆状病毒BV-pdCap。间接免疫荧光和Western blotting试验结果表明,该重组杆状病毒成功表达了具有良好反应原性的pdCap蛋白。 The Cap gene partially deleted nuclear localization signal sequence （pdCap） of porcine circovirus 2 （PCV2） was cloned into pBACsurf-1 between the upstream gp64 signal sequence and downstresm gp64 mature domain to construct the re- combinant plasmid pBACsurf-l-pdCap. The fusion gene containing pdCap and gp64 was subcloned into pFastBacTM Dual to con- struct the recombinant plasmid pFastBac-pdCap. And then the plasmid was transformed into E. coli DH10Bac competent cells to obtain the recombinant shuttle vector Bacmid-pdCap. Eventually, the recombinant Bacmid-pdCap was transfeeted into Sf9 cells to produce the recombinant baculovirus BV-pdCap mediating by LipofectamineTM 2000. Indirect immunofluorescent and Western blotting assay indicated that the recombinant baeulovirus BV-pdCap could express pdCap protein with significant anti- gencity.

关 键 词： 猪圆环病毒 型 蛋白 杆状病毒表达系统

领　　域： [生物学]