Construction And Biological Characteristics Of RV Expressing BVDV-E2 Recombinant Virus

Bovine viral diarrhea is a febrile and acute infectious disease infected through the digestive tract,respiratory tract and placenta.It is distributed worldwide and is one of the major diseases that seriously threaten the cattle industry.Envelope protein E2 is the most dominant antigenic protein of BVDV and has higher immunogenicity.This project uses rabies virus(RV)vaccine strain LBNSE as the viral vector,inserts the BVDV E2 gene into the LBNSE genome,uses reverse genetic manipulation technology to rescue the recombinant virus RV-DE2,and conducts research on its biological characteristics and immune efficacy.The BVDV E2 gene sequence was optimized by the codon optimization software and the E2 gene fragment was obtained.The E2 gene was transformed into the DE2 fragment by the overlap PCR method,and it was inserted into the LBNSE vector by restriction digestion and ligation to construct the recombinant plasmid RV-DE2.Reverse genetic manipulation technology was used to rescue the recombinant virus RV-DE2,and the recombinant virus was identified by direct immunofluorescence method.And to study the biological characteristics of the recombinant virus RV-DE2.The results are as follows:1.Using the rabies vaccine strain LBNSE as a vector,Bsiw ? and Nhe ? as restriction sites,insert the optimized BVDV E2 gene between the G protein and the L protein to successfully construct the RV-DE2 recombinant plasmid;2.The recombinant plasmid and helper plasmid were co-transfected into BSR cells,and the recombinant virus RV-DE2 was successfully rescued;3.The biological characteristics of the recombinant virus were studied and it was found that the characteristics of the recombinant virus and the parent virus were basically the same.The E2 protein of the RV-DE2 recombinant virus was measured and it was found that the expression level of the recombinant virus E2 protein was higher than that of the BVDV virus.The recombinant virus was immunized to mice After successfully producing neutralizing antibodies against the BVDV Ht 01 strain.