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
in Chinese
| WPRIM
| ID: wpr-328018
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.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Antiviral Agents
/
Pharmacology
/
Swine
/
Virus Replication
/
Recombinant Proteins
/
Enzyme-Linked Immunosorbent Assay
/
Gene Expression
/
Cell Line
/
Blotting, Western
/
Baculoviridae
Limits:
Animals
Language:
Chinese
Journal:
Chinese Journal of Biotechnology
Year:
2007
Type:
Article
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