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HCVNS5B基因片段克隆入BAC TO BACTM 重组杆状病毒表达系统的pFASTHTc载体质粒 ,转化DH10BACTM感受态细菌获得重组的Bacmid质粒 ,将重组Bacmid质粒转染Sf9细胞 ,获得的重组杆状病毒可表达目的蛋白。免疫印迹和体外活性检测表明 ,所表达蛋白为HCVNS5B蛋白 ,具有多聚酶活性。
The HCV NS5B gene fragment was cloned into the pFASTHTc vector plasmid of BAC TO BACTM recombinant baculovirus expression system and transformed into DH10BACTM competent bacteria to obtain recombinant Bacmid plasmid. The recombinant Bacmid plasmid was transfected into Sf9 cells, and the recombinant baculovirus obtained can express the target protein. Western blotting and in vitro activity assay showed that the expressed protein was HCVNS5B protein with the activity of polymerase.