Expression of HIV-2 gag gene in Pichia pastoris

Objective: To cany out the secretive express of HW-2 gag protein for developing HTV-2 gag protein vaccine which is against HIV infection.Methods: Recombinant expression plasmid pPIC9-gag was constructed by insterting of HTV-2 gag into yeast expression vector pPIC-9,then transformed into GS115 cells.Positive clones were selected with MD/MM plates and confirmed by PCR.Several clones were incubated in MBGY media and induced by 0.5% methanol in BMMY media.The expression product HTV-2 gag protein was analyzed by SDS-PAGE and confirmed by Western blot.Results:The HTV-2 gag protein could be induced with methanol in Pichia pastoris.The molecular weight of the expressed protein, analyzed by SDS-PAGE,was 55 kD.The Western blot result showed that the expressed protein could be detected by HTV-2 gag specific antibody. Conclusion:The recombinant plasmid HTV-2 gag could be induced to express the HTV-2 gag protein by methanol in methanol-trophic yeast expression system.