Cloning and Prokaryotic Expression of HIV-1 Vif Gene

Objective To clone the sequence of coding region of HIV-1 Vif(virus infectivity factor) gene and express in prokaryotic cells. Methods Amplify Vif gene by PCR and clone to prokaryotic expression vector pMRI. Transform the constructed recombinant plasmid pMRI-Vif to E.coli BL21(DE3) for expression under induction of IPTG. Purify the expressed protein by Ni2+-NTA affinity chromatography and identify by SDS-PAGE and Western blot. Results The restriction map of pMRI-Vif showed the target gene fragment at a length of about 600 bp. Sequencing result proved that the recombinant plasmid was successfully constructed. The expressed Vif protein reached a purity of more than 85% and a protein concentration of about 1 g/L. The purified protein showed good antigen specificity. Conclusion Soluble Vif protein was successfully expressed in E. coli.