鸡传染性贫血病毒VP1、VP2基因在杆状病毒表达系统中的表达

Expression of VP1 and VP2 genes of chicken anemia virus by baculovirus expression systems

将鸡传染性贫血病毒M990 5株VP1、VP2基因分别或同时克隆到杆状病毒转移载体pFastBacDUAL中 ,然后转化到DH10BAC感受态细胞中与Bacmid杆状病毒穿梭载体进行转座重组 ,最后将重组子转染Sf9昆虫细胞 ,得到分别或同时含VP1、VP2基因的重组杆状病毒rBacVP1、rBacVP2及rBacVP1_2。PCR扩增结果证实VP1、VP2基因重组到杆状病毒基因组中 ;SDS_PAGE电泳分析和间接免疫荧光试验结果表明VP1、VP2基因在重组病毒中得到了表达。

Chicken anemia virus strain M9905 was isolated in Mudanjiang region of Heilongjiang province of China in 1999. It's VP1 and VP2 genes encoded by open reading frame (ORF) 1 and 2 were cloned and inserted into the baculovirus transfer vector pFastBacDUAL and then recombined with the baculovirus shuttle vector Bacmid by transforming DH10BAC competent cells. Finally, Sf9 cells were transfected by recombinant Bacmid DNAs and three recombinant viruses that respectively contained VP1,VP2 or VP1+VP2 gene(s) were obtained. PCR amplification confirmed that VP1 and VP2 genes had inserted into the baculovirus genome. Indirect immunofluorescence assay and SDS-PAGE analysis showed that vp1 and vp2 has been expressed in Sf9 cells infected with the recombinant baculoviruses.