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Chinese Journal of Biotechnology ; (12): 386-391, 2007.
Artigo em Chinês | WPRIM | ID: wpr-328018

RESUMO

The full-length porcine interferon gamma(PoIFN-gamma) cDNA, including the secretion signal peptide coding region was recloned into honor plasmid pFastBac 1 of Bac-To-Bac Baculovirus Expression System. These recombinant plasmids, pFastBac -PoIFN-gamma, were transformed into DH(10Bac) host bacteria to get recombinant shuttle plasmids, rBacmid-PoIFN-gamma. Recombinant baculovirus, rBac-PoIFN-gamma, was generated for expressing PoIFN-gamma, by transfecting rBacmid-PoIFN-gamma with Cellfectin Reagent into sf9 insect cells. The expression of PoIFN-gamma in insect cells was confirmed by Western Blot, indirect immunofluorescence assay and indirect ELISA. The antiviral activity assay shows that PoIFN-gamma expressed by the rBac-PoIFN-gamma can efficiently inhibit the replication of the recombinant Vesicular Stomatitis Virus expressing green fluorescence protein in PK-15 cells. The antiviral activity of PoIFN-gamma can be specifically blocked by anti-PoIFN-gamma mouse serum. The antiviral titer of culture supernatant of insect cells infected by rBac-PoIFN-gamma is 2 x 10(4) IU/mL. The results demonstrat that the rBac-PoIFN-gamma can express rPoIFN-gamma efficiently and rPoIFN-gamma has high antiviral activity.


Assuntos
Animais , Camundongos , Antivirais , Metabolismo , Farmacologia , Baculoviridae , Genética , Western Blotting , Linhagem Celular , Ensaio de Imunoadsorção Enzimática , Técnica Indireta de Fluorescência para Anticorpo , Expressão Gênica , Proteínas de Fluorescência Verde , Genética , Metabolismo , Soros Imunes , Alergia e Imunologia , Interferon gama , Genética , Alergia e Imunologia , Metabolismo , Farmacologia , Camundongos Endogâmicos BALB C , Microscopia de Fluorescência , Proteínas Recombinantes , Spodoptera , Suínos , Vesiculovirus , Genética , Replicação Viral
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