[Expression of bovine interferon gamma in recombinant baculovirus and determination of its antiviral activity].
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.
Qin
Li-ting
LT
Key Laboratory of National Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agriculture Science, Harbin 150001, China. qinliting@yahoo.com.cn
Wang
Xi-jun
XJ
Hu
Sen
S
Liu
Si-dang
SD
Bu
Zhi-gao
ZG
D000818:Animals; D000998:Antiviral Agents / Q000378:metabolism / Q000494:pharmacology*; D016367:Baculoviridae / Q000235:genetics* / Q000378:metabolism; D002417:Cattle; D002460:Cell Line; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D007371:Interferon-gamma / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology*; D011994:Recombinant Proteins; D014721:Vesicular stomatitis Indiana virus / Q000187:drug effects
D000998:Antiviral Agents; D011994:Recombinant Proteins; D007371:Interferon-gamma