Expression of HIV-1 gp 120 in methylotrophic Pichia Pastoris

Aim To express HIV-1 protein gp 120 in Pichia Pastoris.Methods A long fragment(1 419 bp) and a short fragment(417 bp) of gp 120 gene were amplified from HIV-1 international standard strain pHXB2-gp 160,and were sub-cloned into eukaryotic expression vector pPICZαA and pPICZB.The constructed plasmid was transformed into yeast GS115 by electroporation.The recombinant transformants were selected by YPDS plates,and PCR was used to test the insert.The expression in yeast was induced by the addition of methanol and was analyzed by SDS-PAGE and Western blot.Results The gp 120 short peptide was expressed in GS 115.At 24 hours induction time,its expression level and antigenecity are the highest.The expressed product was truncated and showed excellent antigenic specialty.The gp120 long peptide was not expressed by GS115.Conclusion Gene optimization should be performed to express successfully the gp 120 long DNA fragment in Pichia Pastoris.the research is useful for the production of diagnostic reagents and genetically engineered vaccine of HIV-1.