Construction,expression and purification of HIV-1 p24-gp41 fusion protein in E.coli / 中华传染病杂志

ABSTRACT

Objective To construct recombinant plasmid with p24 and gp41 gene, express fusion protein in E.coli. Methods To design primer with restriction endonuclease position,and amplify p24 and gp41 by RT-PCR, link both into pMD18-T vector.To choose correct clone with target gene.Then p24 fragment will be cleaved and linked into pMD18-T vector within gp41 gene. Both post-linked gene will be cleaved and linked into pET21a vector. The vector will be transformed into E.coli. And protein is highly effective expressed in E coli. Western blotting proved that the expressed product could react with 6?his antibody. Result Fusion protein p24-gp41 is highly effective expressed in E.coli. Conclusion Fusion protein p24-gp41 is highly effective expressed in E.coli in pET21a vector.