Study On The Effect Of Vimentin In BVDV Infect MDBK

Bovine viral diarrhea(BVD)is one of the most important infectious diseases that seriously harms the cattle industry.Studying the infection mechanism of the virus on the host contributes to the prevention and control of the disease.In our previous study,four interacting proteins that may interact with BVDV and have the most follow-up value were screened for vimentin,α-actinin,actin and tubulin.A large number of studies have shown that vimentin plays an important role in the infection of many viruses.This study is to determine the role of vimentin in the infection of MDBK cells by BVDV.This study has important theoretical value for understanding the mechanism of BVDV infection.This study first determined whether BVDV infection the expression of vimentin in MDBK cells.The transcription level of vimentin in MDBK cells after BVDV infection was detected by real-time quantitative PCR(q RT-PCR),and the level of vimentin was detected by Western blot.The results showed that in the early stage of BVDV infection,the transcription and protein content of vimentin in MDBK cells were significantly reduced.Laser confocal detection confirmed co-localization of vimentin with BVDV structural protein E2.It is speculated that it interacted with vimentin during BVDV infection.Second,in order to determine the effect of disruption and knockdown of vimentin on viral replication in BVDV-infected MDBK.Using short hairpin RNA-mediated RNA interference technology,design and construct a specific sh RNA plasmid targeting the vimentin,use sh RNA to knock down the expression of vimentin in MDBK cells,destroy the vimentin structure with ACY,and then infect BVDV to determine destruction and knock The effect of low waveform proteins on viral replication.The results showed that three specific sh RNA plasmids targeting the vimentin were successfully constructed,which knocked down the expression of endogenous vimentin of MDBK.When the vimentin is knocked down or destroyed on MDBK cells,it affects the replication of BVDV and reduces the virus titer.To further investigate the effect of recombinant vimentin and vimentin monoclonal antibody on viral replication in BVDV-infected MDBK.The vimentin gene was amplified by PCR,and the Vim-p ET-28 a expression plasmid was constructed to induce expression,and the target protein was purified by a nickel column.MDBK cells were coated with recombinant vimentin and vimentin monoclonal antibody to detect viral replication when BVDV was infected with MDBK.The results showed that the prokaryotic expressed vimentin was successfully obtained.Both recombinant vimentin and vimentin monoclonal antibody blockade have an effect on earlyreplication of BVDV,significantly reducing viral titers.Taken together,BVDV infection inhibits the transcription and expression of vimentin in MDBK.After vimentin is knocked down or destroyed,the level of replication of BVDV decreases.After blocking by vimentin and vimentin,both of them affected the replication of BVDV and reduced the virus titer.Vimentin is associated with viral replication when BVDV is infected with MDBK cells.This study helps to reveal the infection mechanism of BVDV and lays a foundation for the prevention and control of BVDV.