Expression of rabbit hemorrhagic disease virus VP60 protein with B cell epitope deletion in N terminus and its immunogenicity in rabbit
This study used the NTA region of the VP60 protein recognized by A3C monoclonal antibody as a recogni-tion marker,designed a series of gene sequences encoding overlapping peptides in the VP60 NTA region,and connected them to the pGEX-4T-1 vector to determine the precise epitopes recognized by A3C monoclonal antibody.Then recombinant plasmid Bacmid-VP60△A3C was con-structed.Bacmid-VP60△A3C was transfected into Sf9 in-sect cells,and recombinant baculovirus rBac-VP60△A3C was obtained.The identification results of RT-PCR,HA,IFA,and Western blot showed that the recombinant protein VP60△A3C was highly expressed in Sf9 cells.Electron micros-copy observation showed that its morphology and structure were similar to those of rabbit hemorrhagic disease virus VP60 protein.The 2-month-old RHDV serum negative New Zealand rabbits were immunized with recombinant protein VP60△A3C at a rate of 200 μg per rabbit,and New Zealand rabbits were challenged with rabbit hemorrhagic disease virus WF strain after 14 days.The results showed that there were no deaths in the immune group,while all deaths occurred in the control group.The results of this study lay the foundation for the development of epitope-deleted subunit vaccine of rabbit hemorrhagic disease.
NETL
NSTL
万方数据
维普
