Inhibition Of Chicken TRIM62 On The Replication Of Reticuloendotheliosis Virus

Avian Reticuloendotheliosis virus(REV)is a retrovirus that causes severe immunosuppression and tumor in poultry.Currently,there are no effective vaccines or drugs for prevention and control,causing serious damage to the poultry industry.TRIM62(tripartite motif containing 62)is a host defensive protein that plays an important role in immune-related signaling pathways and antiviral immune responses.It is a new natural immune regulatory factor.In our previous study,we conducted a proteomic analysis of DF-1 cells infected with REV.We found that the expression of TRIM62 was inversely related to the replication of virus.Therefore,it was hypothesized that chicken TRIM62 may inhibit the replication of REV.Studies have reported that human TRIM62 has the effect of inhibiting HIV replication of retroviruses.The inhibitory effect of chicken TRIM62 on retrovirus replication has not been reported.Based on the data of previous experiments and the biological functions of TRIM62,we designed this topic to explore the role of TRIM62 antiretroviral REV,providing a preliminary basis for further study of its anti-retroviral mechanism.Firstly,tissue RNA was extracted from DF-1 cells,and the chicken TRIM62 protein gene was obtained by PCR amplification.TRIM62 protein was obtained after prokaryotic expression and protein purification.The transfected TRIV62 protein and lentiviral recombinant vector were transfected into REV-infected DF-1 cells.The anti-REV effect was detected by fluorescence quantitative PCR and Western Blot.The purified TRIM62 protein was used to immunize New Zealand white rabbits to prepare chicken TRIM62 polyclonal antibody.The distribution and dynamic expression of TRIM62 in chicken tissues were detected by immunohistochemistry.The qRT-PCR method was used to detect the level of dynamic transcription of TRIM62 in chicken tissues.The results showed that the TRIM62 protein expressed and purified was a single band;TRIM62 could significantly inhibit the replication of REV in DF-1 cells,and its inhibitory effect was positively correlated with the expression level.The results of immunohistochemistry showed that TRIM62 is widely distributed in various tissues and located in the cytoplasm.The expression of TRIM62 intissues is related to the tissue tropism of REV.REV infection affects the transcription and expression levels of TRIM62,and the affects is differently in different stages of virus infection.In this study,a specific TRIM62 polyclonal antibody was prepared,and it was clarified that chicken TRIM62 protein can effectively inhibit the replication of REV;TRIM62 is widely distributed in various tissues and located in the cytoplasm.The expression of TRIM62 in tissues is related to the tissue tropism of REV.The study lays the foundation for the study of the late TRIM62 anti-REV mechanism.