Production and evaluating the properties of HIV-1-Nef-MPER-V3 fusion protein harboring IMT-P8 cell penetrating peptide.
2020
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.
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