| Title | Secretory expression of porcine interferon-gamma in baculovirus using HBM signal peptide and its inhibition activity on the replication of porcine reproductive and respiratory syndrome virus. | | Author(s) | Wang YB, Wang ZY, Chen HY, Cui BA, Wang YB, Zhang HY, Wang R | | Institution | Key Laboratory of Animal-derived Food Safety of Henan Province, College of Husbandry and Veterinary, Henan Agricultural University, Zhengzhou, Henan 450002, China. | | Source | Vet Immunol Immunopathol 2009 Jun 6. | | Abstract | The gene sequence encoding mature porcine interferon-gamma (PoIFN-gamma) fused with a C-terminal 6x histidine tag was cloned into the baculovirus pFastBac Dual vector of the Bac-to-Bac Baculovirus expression system under the control of PH promoter. The authentic signal sequence of porcine interferon-gamma was substituted with the honeybee melittin (HBM) signal sequence, and expressed in insect cells. The recombinant proteins were detected by SDS-PAGE and immunofluorescence assay. The nickel affinity column purified recombinant porcine interferon-gamma with HBM signal peptide (rPoIFN-gammaH) was shown to be a 19kDa protein as confirmed by Western blot analysis. The recombinant PoIFN-gammaH was shown to have cytokine activity, inhibiting the cytopathic effect of vesicular stomatitis virus (VSV) in PK-15 cells at about 1.07x10(6)U/mL. The 2(-7) dilution of the rPoIFN-gammaH in culture supernatant protected the MARC-145 cells from the cytopathic effect caused by 100TCID(50) of porcine reproductive and respiratory syndrome virus. | | Language | ENG | | Pub Type(s) | JOURNAL ARTICLE
| | PubMed ID | 19556014 |
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