2010•Zhongguo yufang shouyi xuebaoRequires access
Establishment of BHK-21 cell lines stably expressing T7 RNAP.
Xin Li, Yang ShaoHua, Hongmei Wang, Xiao Liu, Jianming Wu, Yun‐Dong Gao, Liqun Wang, Jifeng Zhong, Hongbin He
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Abstract
To establish the cell line with stable expression of T7 RNA polymerase gene,T7 RNAP gene was amplified from E.coli BL21(DE3) and inserted into pcDNA3.1(+) vector to construct recombinant plasmid pcDNA-T7RNAP.The BHK-21 cell line stably expressing T7 RNAP was generated by pcDNA-T7RNAP transfection and the positive cell clones were obtained after continually screening using G418.The expressions of T7 RNAP gene were confirmed by RT-PCR and western blot.The cell line stably expressing the T7 RNAP gene could provide a platform for rescue RNA recombinant virus in vitro via reverse genetic technique.