Abstract

The gene sequence encoding mature porcine interferon-gamma (PoIFN-γ) fused with a C-terminal 6× 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-γH) was shown to be a 19 kDa protein as confirmed by Western blot analysis. The recombinant PoIFN-γH was shown to have cytokine activity, inhibiting the cytopathic effect of vesicular stomatitis virus (VSV) in PK-15 cells at about 1.07 × 10 6 U/mL. The 2 −7 dilution of the rPoIFN-γH in culture supernatant protected the MARC-145 cells from the cytopathic effect caused by 100TCID 50 of porcine reproductive and respiratory syndrome virus.

Full Text
Paper version not known

Talk to us

Join us for a 30 min session where you can share your feedback and ask us any queries you have

Schedule a call