Expression and Characterization of Two DNA Constructs Derived from HIV-1-vifin Escherichia coli and Mammalian Cells

Background: acquired immunodeficiency syndrome (hiv/aids) is still a major global concern and no effective therapeutic vaccine has been produced to prevent the problem. among hiv-1 proteins, vif as a basic cytoplasmic protein of hiv-1 is involved in late stages of viral generation and plays important role in hiv-1 virion replication. it also increases the stability of virion cores, which probably inhibits early degradation of viral entry. therefore, it seems rational to apply this protein as a vaccine based on its impact on hiv-1 life cycle. this study aimed at cloning, expression and production of vif protein as an hiv-1 vaccine candidate. methods: in this study, vif sequence was amplified from pln4-3 plasmid including hiv-1 vif gene and then cloned in pet23a to generate the recombinant plasmids of pet23a/vif with hexahistidine tags. bl21 competent cells were transformed to obtain the protein of interest. ni-nta column was used to purify the protein of interest and western blotting confirmed vif protein using anti-his tag antibody. in order to express the gene of interest in eukaryotic cells, vif was sub-cloned into pegfp plasmids and hek 293-t cells were transfected. flow cytometry was then applied to evaluate gfp expression. results: vif protein was expressed in bl21)de3) strain and identified as a23 kda band in sds-page and confirmed by anti-his antibody in western blotting. the purified protein concentration was 173.3 μg/ml using bradford assay. hek-293t cells were successfully transfected by recombinant pegfp plasmids and flow cytometry confirmed the cell transfection. conclusion: vif protein can be expressed in mammalian cells and may be a proper protein subunit vaccine candidate against hiv-1.