Construction And Expression Of The Recombinant Fowlpox Virus Expressing The E0 Antigen Of Classical Swine Fever Virus Shimen Strain And Animal Immunity Experiment

Classical swine fever(CSF) caused by Classical swine fever virus(CSFV) was highly fetal infection disease ,and was harmful in the global swine breeding.CSF was classified as class A infectious disease by OIE.CSFV was RNA virus with envelope.inclusive of three kinds of envelope protein E0(Erns),El and E2, the production of antibody induced by CSFV was relative to E0 ,the sequency coding E0 protein was conservative.Therefore ,it could also be used as a subunit vaccine. Inactivate vaccine and weaken live vaccine were efficient in preventin from CSF. But the infection of vaccine could be discriminated with natual infection. Labelled vaccine could be the solution to the problem mentioned above.In this paper, CSFV E0 gene was transferred into Fowlpox virus in order to obtain CSFV live vector vaccine. This paper was mainly about three parts below:1.The construction of recombinat migration vector,a series of molecular biological methods were performed,E0 gene was transferred into migration vector FPV-pSY named as FPV-pSY-E0.The result of enzyme digestion and PCR identification showed the construction was successful.the recombinat migration vector was purified for transfection with Wizard purifiction Plasmid kit.2.The screening and purification of recombinant Fowlpox virus. FPV-pSY-E0 purified was transinfected into Chicken embryo fibroblast (CEF) infected by Fowlpox virus,FV282-E0 was purified to obtain the stable recombinant Fowlpox virus expressing E0 antigen by blue plaque screening for 10 times PCR amplification,SDS-PAGE and Western blot were performed to identify the recombinant Fowlpox virus,the results showed that CSFV E0 antigen was expressed successfully in this recombinant Fowlpox virus,and could provide important sample to next experiment.3. The immunity experiment was separated into two steps.Firstly,the Balb/c mice were injected for 3 times with the recombinant Fowlpox virus, the titer of CSFV antibody was 1:4096;Secondly, the swine immunity experiment was performed to evaluate the protection effect of recombinant Fowlpox virus.the pretction ratio was 75%.the animal experiment indicated that high titer of CSFV antibody could be induced and the pigs could be protected well by the recombinant Fowlpox virus expressing E0 antige.This paper could be the foundation for prevention of CSF.