Coexpression of Vif and ElonginC Proteins of HIV-1 in E. coli

Objective To coexpress Vif and ElonginC proteins of HIV-1 in E. coli. Methods Amplify the full-length of DNA encoding ElonginC protein by PCR and clone into plasmid pMDT-easy. Digest the constructed recombinant plasmid with NdeⅠ/ BamHⅠ, and insert the obtained target gene into plasmid pRSETB to construct recombinant plasmid pRSETB-ElonginC. Transform recombinant plasmid pMRI-Vif constructed previously to E. coli BL21(DE3), and prepare the obtained recombinant E. coli into competent cells. Transform recombinant plasmid pRSETB-ElonginC to the prepared competent cells for expression under induction of 1 mmol / L IPTG. Identify the expressed product by SDS-PAGE and Western blot. Results Two gene fragments, at lengths of about 3 000 and about 400 bp respectively, were obtained by digestion of pRSETB-ElonginC with Nde Ⅰ/ BamHⅠ, which proved that the recombinant plasmid was constructed correctly. SDS-PAGE and Western blot proved that both Vif and ElonginC proteins were expressed and showed good antigenic specificity. Conclusion Vif and ElonginC proteins were coexpressed in E. coli.