Expression of HIV-1 Gag Protein (Chinese Strain) in Pichia Pastoris

[Abstract] Objective To construct a recombinant plasmid pPICGAG for the expression of whole length of HIV - 1 Gag gene in Pichia pastoris . Methods The plasmid pKSGAG containing the whole length of HIV - 1 Gag gene was digested with Not I and Xhol and cloned into expression vector pPIC9. The constructed plasmid pPICGAG was linarized with Sac I and transformed to Pichia pastoris GS 115.The positive transforma-nts were identified by PCR and expressed in BMGY and BMMY media.The expressed products were analyzed by SDS - PAGE. Results The integration rate of transformant was 72.7% . SDS - PAGE showed a relative molecular weight of expressed protein of about 55000, and Western blot showed specific reaction of it with McAb. Conclusion The HIV - 1 Gag protein with good reactogenicity and specificity was successfully expressed in Pichia pastoris.