Expression of HIV-2 gp105-gag truncated gene in Pichia pastoris
Objective To carry out the secretive express of HIV 2 ROD tgp105 gag protein and the optimization of expression conditions. Methods HIV 2 gp105 truncated gen (tgp105) was obtained by PCR amplification and was cloned into a secreting expression vector pPIC 9. Recombinant expression vector pPIC9 tgp105 gag was constructed by inserting HIV 2 gag into pPIC9 tgp105 recombinant plasmid, then transformed into GS115 cells. Positive clones were selected with MD/MM plates and confirmed by PCR. Several clones were incubated in BMGY media and induced by 0.5% methanol in BMMY media. The expression product HIV 2 tgp105 gag protein was analyzed by SDS PAGE and confirmed by Western blot. Results The tgp105 gag protein was secreted into media. The molecular weight of the expressed protein, as analyzed by SDS PAGE, was 140kD. The Western blot result showed that the expressed protein could be detected by HIV 2 specific antibody. Conclusion The recombinant plasmid HIV 2 ROD tgp105 gag was successfully expressed in Pichia pastoris and the expressed protein has a good antigen specificity. [