Expression of bovine interferon gamma in recombinant baculovirus and determination of its antiviral activity
牛γ-干扰素在重组杆状病毒中的表达及其抗病毒活性的测定

The full-length bovine interferon gamma (BoIFN-gamma) cDNA, including the secretion signal peptide coding region was recloned into baculovirus honor vectors pFastBac 1 of Bac-To-Bac system. These recombinant plasmids, pFastBac 1-BoIFN-gamma, were transformed into DH(10Bac) host bacteria to get recombinant shuttle plasmids, rBacmid-BoIFN-gamma. Recombinant baculovirus, rBac-BoIFN-gamma, was generated for expressing BoIFN-gamma, by transfecting recombinant Bacmid-BoIFN-gamma with Cellfectin Reagen into sf9 insect cells. BoIFN-gamma efficiently expressed by recombinant baculovirus in sf9 cells was testified by indirect immunofluorescence assay and indirect ELISA with monoclonal antibody against Bovine interferon-gamma. Furthermore, VSV * GFP, recombinant Vesicular Stomatitis Virus expressing green fluorescence protein and MDBK were used to determine the anti-viral activity of rBoIFN-gamma. The result shows rBoIFN-gamma could inhibit the replication of the VSV * GFP in MDBK cells and the antiviral activity of supernatant was 2 x 10(5) IU/mL. The antiviral activity of rBoIFN-gamma could be blocked by anti-BoIFN-gamma mouse serum. The results demonstrated that the recombinant baculovirus could express BoIFN-gamma efficiently and rBoIFN-gamma had high antiviral activity.