• Expression of the HIV-1 Nef-Vif fusion protein in E. coli strain
  • Fateme shahhosaini,1,* Azam Bolhassani,2 Neda Mousavi-Niri,3
    1. Department of Biotechnology, Faculty of Advanced Science and Technology, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran
    2. Department of Hepatitis and AIDS, Pasteur Institute of Iran, Tehran, Iran
    3. Department of Biotechnology, Faculty of Advanced Science and Technology, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran


  • Introduction: The development of an effective vaccine against the human immunodeficiency virus (HIV)-1 is a global health priority. Fusion recombinant vaccines have several potential benefits which could be applied in the case of mutable divergent pathogens. HIV-1 is a complex retrovirus containing 15 different proteins which play key roles in the virus's pathogenicity. Since accessory proteins of HIV-1are critical throughout the infection, a fusion recombinant protein containing the full length of HIV-1 Nef and Vif was expressed in E. coli Rosetta strain.
  • Methods: At first, cloning of the Nef-Vif fusion gene was performed in the pET24a (+) expression vector. Then, the recombinant Nef-Vif protein was expressed in Rosetta E. coli bacteria using IPTG inducer in different conditions. Finally, the protein expression was confirmed by SDS-PAGE and western blotting.
  • Results: The recombinant pET24a (+)-Nef-Vif vector was confirmed by enzymatic digestion and PCR demonstrating a clear band of ~ 1200bp related to the Nef-Vif fusion gene conducting agarose gel electrophoresis. SDS-PAGE and Western blotting confirmed the expressed protein as a clear band of ~50 kDa at 16 h after induction with 1mM IPTG.
  • Conclusion: Considering the important roles of Nef and Vif in virus infectivity, the construction of Nef-Vif fusion gene was performed in a prokaryotic expression vector to express the Nef-Vif fusion protein in E.coli strain under certain conditions. This protein will be purified in progress to develop a protein-based HIV-1 vaccine candidate in future.
  • Keywords: AIDS, HIV, Nef, Vif, Recombinant protein