1.Construction and identification of a vector inserted with gene of T7 RNA Polymerase
Hong-Hui SHEN ; Bing-Ke BAI ; Hao-Dong LIU ; Sheng-Dong LUO ; Yan HU ; Jun HOU ; Zhi-Fie WANG ; Wei KONG ; Yi-Dan BAO ; Pan-Yong MAO
Chinese Journal of Experimental and Clinical Virology 2011;25(2):146-148
Objective To develop a system to rescue virus by intracellular expression of T7 RNA Polymerase. Methods The gene of T7 RNA Polymerase was amplified and cloned to VR1012 by molecular biological technology. The expression plasmid VR-1a was then identified. VR-1a and EV71 infectious plasmid were co-transfected in Veto cell. CPE was observed and viral gene viral antigen were detected.Results The gene of T7 RNA Polymerase was successfully cloned into vector VR1012. Vere cell developed to CPE after being transfected VR-1a and EV71 infectious plasmid. EV71 gene was amplified by RT-PCR from the culture. EV71 antigen was also detected by ELISA. Conclusion The method can be used to rescue virus. It could apply to immunologic research of EV71 DNA vaccine.
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