Comparative study of three different transfection methods to rescue the virus

Objective Comparing three different transfection methods to explore the best strategy of rescuing virus.Methods Three transfection methods include:① EV71 infectious plasmids including T7 RNA polymerase promote and T7 RNA polymerase eukaryotic expression plasmid were co-transfected in Vero cell.② Full-length PCR products of EV71 including T7 RNA polymerase promote and VR-1a were cotransfected in Vero cell.③ Virus RNA of EV71 was transcripted by T7 ribomaxTM epress large scale RNA production system was transfected in Vero cell directly.Respectively observe the time of producing CPE after transfecting.Viral gene and viral antigen were detected by RT-PCR and WB.Results Vero cell can be successfully transfected by three genetic patters.Transfection effect ③ is better than that of ②,② is better than that of①.Conclusion Three genetic patters can be used to rescue virus.The best Effect was viral RNA transfection directly. Key words: RNA;  Enterovirus;  Transfection