Generation Of Recombinant Baculovirus Expressing VP2 Protein Of Mink Enteritis Virus And Analysis Of Its Immunogenicity In A Rabbit Model

Mink viral enteritis is highly contagious and lethal disease of minks caused by mink enteritis virus(MEV).The key feature of this disease is diarrhea. The Incidence and mortality is very high. It is generally acknowledged as one of the disease, which make breeding mink company lose very much. As the other disease, vaccination has been an important measure to prevent MEV.Virus-like particles is s a self-assembly vacant capsid structure made up by one or a few structural protein of virus. Virus-like particles is like immaturity virion. Because it is lace of infectious nucleic acid and regulatory proteins, Virus-like particles has no ability of replication or infection. But VLPs could be intook and transported by cells. It could provocation full immunity. Utilizing the expression system of baculoviruses, this paper has enunciated the expressing of MEV vp2 gene and developed an experimental immunity research.This study is basis on the MEV VP2 protein expressed with baculoviruses could assemble virus-like particles itself. And the virus-like particles still have Immunogenicity.This study generate a recombinant baculovirus which could express MEV VP2 protein. This study has established groundworks for the research of the MEV subunit vaccines.SMPV-11 is one of strains of MEV. SMPV-11 VP2 gene was amplified by PCR method and then cloned into pMD18-T simple vetcor.Sequencing the VP2 gene showed that the VP2 gene's antigenic type is MEV-1 type which is most popular in China. The VP2 gene was inserted into pFastBacâ… vector, then transferred the recombinant plasmid pFastBacâ… -VP2 into DH10Bac to obtain recombinant Bacmid. Subsequently, the recombinant Bacmid was transfected into Sf9 cells. Electron microscope shows that rBacMEV-VP2 could express VP2 protein,and it could assemble VLPs itself.Identified by Western-blotting and indirect immunofluorescent assay , the expressed recombinant protein shows well reactivity with MEV monoclonal antibody.The expressed recombinant protein was centrifugated and purificated.Mix the recombinant protein with alhydrogel as vaccines. Inoculate oryctolagus cuniculus with 100ug,500ug expressed recombinant protein.Two weeks later , the HI antibody could be detected.Three weeks after inoculating, the HI antibody value is highest, as 1:108.8,1:204.8.The study suggested that the expressed recombinant protein is as well as MEV inactivation vaccines.