Expression of Human Immunodeficiency Virus-1 gp120 and gp41 Gene Synthesized in Pichia pastoris

Objective To synthesize HIV-1 envelope gene gp120 and gp41 which was suitable for yeast protein expression and to express synthetic genes in Pichia pastoris.Methods Using synonymous codons preferred by yeast usage on protein expression to replace some native codons of wild-type HIV genes,the synthetic gp120 and gp41genes were achieved by a recursive PCR(rPCR).The genes synthesized were inserted into the yeast expression vector pPICZɑA.The recombinant plasmids were transformed into GS115 yeast by electroporation.The yeast transformants induced by methanol expressed gp120 and gp41 protein.Results The restriction analysis and DNA sequencing confirmed that the synthetic genes were inserted to yeast pPICZɑA in correct orientation.SDS-PAGE and immunoblotting indicated that the secreted forms of gp120 and gp41 were expressed by yeast transformants.Conclusion The secreted forms of recombinant gp120 and gp41 were expressed efficiently in Pichia pastoris by using synthetic genes.