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

To develop a system to rescue virus by intracellular expression of T7 RNA Polymerase.

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.

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.

The method can be used to rescue virus. It could apply to immunologic research of EV71 DNA vaccine.