利用嵌合内含子增强HIV-1 Vif蛋白的体外表达

OBJECTIVE Construct eukaryotic expression plasmids of HIV-1 vif gene with chimeric intron to improve the expression of HIV-1 Vif in vitro for better study of its biological function. METHODS Adding an intron between the promoter of the vector and open reading frame (ORF) of vif to generate Vif-expressing plasmid pcDNA3.1-Intron-Vif-HA. The western blotting experiment was performed to compare the different expression levels between pcDNA3.1-intron-Vif-HA and pcDNA3.1-vif-HA. Co-transfecting with APOBEC3G-expressing plasmid, the ability of Vif to degrade APOBEC3G was detected to conform the normal function of Vif. RESULTS The western blotting result showed that the expression of Vif in pcDNA3.1-intron-vif-HA is much higher than it in control plasmid (pcDNA3.1-vif-HA). And Vif expressed by the plasmid with intron could degrade APOBEC3G very well in vitro. CONCLUSIONS Eukaryotic expression plasmid of HIV-1 vif gene with chimeric intron enhances the expression of Vif significantly, and the protein expressed by this plasmid presents the normal function of Vif.