The Construction and Rescue of Epitope Mutant Strain of Classical Swine Fever Virus C Strain
An infectious clone was constructed by mutating the WH303 site of CSFV C strain to provide candidate strains for the study of marker vaccine of CSFV C strain.In this study,CSFV C strain was used as the research material,and WH303 site was mutated by overlapping PCR method,and then cs-full and CS-full-flag were cloned into the full-length infectious clones of CS-FV C strain.In vitro transcription RNA,and then transfected into PK15 cells,continuous pas-sage of cells to complete the cell rescue and proliferation.The results showed that the specific re-striction endonuclease digestion reaction verified the successful construction of infectious clone.RT-PCR and peroxidase monolayer cell test detection indicated that the virus had been rescued successfully.The virus can still be detected in the 17th generation of virus cell,indicating that the virus can be stably passaged.In summary,MC and MC-flag of C strain WH303 mutated in-fectious clones were constructed successfully in this study,MC strain was saved successfully,and the stable virus strain was obtained.
万方数据
维普
