Expression of porcine gamma-interferon in recombinant baculovirus and determination of its antiviral activity / 生物工程学报
Chinese Journal of Biotechnology
; (12): 386-391, 2007.
Article
em Zh
| WPRIM
| ID: wpr-328018
Biblioteca responsável:
WPRO
ABSTRACT
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.
Texto completo:
1
Índice:
WPRIM
Assunto principal:
Antivirais
/
Farmacologia
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Suínos
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Replicação Viral
/
Proteínas Recombinantes
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Ensaio de Imunoadsorção Enzimática
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Expressão Gênica
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Linhagem Celular
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Western Blotting
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Baculoviridae
Limite:
Animals
Idioma:
Zh
Revista:
Chinese Journal of Biotechnology
Ano de publicação:
2007
Tipo de documento:
Article