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Objective To measure the effect of duck viperin protein on the proliferation of duck Tembusu virus (DTMUV). Methods We analyzed the duck viperin gene using a bioinformatics. Plasmids pGEX-6P-viperin and pEGFP-N1-viperin were constructed and transformed into Rosseta and transfected into BHK-21 cells, respectively. BHK-21 cells transfected with plasmid pEGFP-N1-viperin and empty vector were infected with DTMUV. The content of DTMUV in cell precipitation and supernatant was analyzed by real-time fluorescent quantitative PCR. Finally, the effect of viperin on DTMUV proliferation and the binding proteins captured by EGFP monoclonal antibody were evaluated using mass spectrometry. Results Significant differences were observed between the expression levels of viperin gene in duck compared to other species. Duck viperin was found to inhibit the budding of DTMUV in BHK-21 cells. Further, we identified six proteins that might be involved in the inhibition of the proliferation and budding of DTMUV, possibly indicative of the viperin anti-Tembusu virus pathway in ducks. Conclusion Expression patterns of duck viperin reveal how the budding of duck Tembusu virus is inhibited.

Objective To obtain the envelope protein B2L of orf virus(ORFV) and prepare highly specific polyclonal antibody against B2L. Methods The B2L gene was amplified by PCR, and subcloned into the vectors pET-32a and p3×FLAG-CMV-14, pET-32a-B2L followed by expression of B2L protein in E.coli, and induction of the target protein by IPTG, purification by urea solution and identification via Western blot analysis. Furthermore, the BALB/c mice were immunized with B2L protein to prepare highly-specific anti-B2L polyclonal antibody. In addition, plasmid p3×FLAG-B2L was transfected into Vero cells and BHK-21 cells which was used to confirm its specificity and antigenicity by indirect immunofluorescence assay(IFA). Results Western blotting demonstrated that the B2L protein had high-level specificity. The results of IFA indicated that the anti-B2L specific polyclonal antibody had specificity and reactivity. Then Immunohistochemistry showed that it could neutralize natural ORFV. Conclusion Highly specific and purified polyclonal antibody against B2L protein of the ORFV was successfully prepared.