[Construction and identification of a vector inserted with gene of T7 RNA polymerase].

ABSTRACT

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 Vero cell. CPE was observed and viral gene viral antigen were detected. RESULTS: The gene of T7 RNA Polymerase was successfully cloned into vector VR1012. Vero 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.