Shekoufa Jahedian, Seyed Mehdi Sadat*, Gholam Reza Javadi and Azam Bolhassani Pages 1 - 9 ( 9 )
Background: Finding a safe and effective vaccine against HIV-1 infection is still a major concern and highly valuable.
Objective: This study aimed to design and produce a recombinant Nef-MPER V3 protein fused with IMT-P8, using E. coli expression system to provide a potential HIV vaccine with high cellular penetrance.
Methods: After synthesizing the DNA sequence of the fusion protein, the construct was inserted into pET-28 expression vector. The recombinant protein expression was induced using 1 mM IPTG and the product was purified through affinity chromatography. Characterization of cellular delivery, toxicity and immunogenicity of the protein was carried out. Results: The recombinant protein was expressed and confirmed by anti-Nef antibody through western blotting. Data analyses showed that the protein has no considerable toxicity effect and has improved the IMT-P8 penetration rate in comparison with a control sample. Moreover, the antigen immunogenicity of the protein induced specific humoral response in mice.
Conclusion: It was concluded that IMT-P8- Nef-MPER-V3 fusion protein has a high penetrance rate into mammalian cell line and low toxicity for potential application as a vaccine against HIV-1.
HIV-1, Recombinant protein, Escherichia coli, Nef-MPER-V3, Cell penetrating peptide, IMT-P8.
Department of Biology, Sciences and Research Branch, Islamic Azad University, Tehran, Department of Hepatitis, AIDS and Blood-borne diseases, Pasteur Institute of Iran. Tehran, Department of Biology, Sciences and Research Branch, Islamic Azad University, Tehran, Department of Hepatitis, AIDS and Blood-borne diseases, Pasteur Institute of Iran. Tehran