Production and Evaluation of the Properties of HIV-1-Nef-MPER-V3 Fusion Protein Harboring IMT-P8 Cell Penetrating Peptide

免疫原性 重组DNA 融合蛋白 抗体 分子生物学 病毒学 抗原 生物 化学 免疫学 生物化学 基因
作者
Shekoufa Jahedian,Seyed Mehdi Sadat,Gholam Reza Javadi,Azam Bolhassani
出处
期刊:Current HIV Research [Bentham Science Publishers]
卷期号:18 (5): 315-323 被引量:3
标识
DOI:10.2174/1570162x18666200612151925
摘要

Finding a safe and effective vaccine for HIV-1 infection is still a major concern.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.After synthesizing the DNA sequence of the fusion protein, the construct was inserted into the 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.The recombinant protein was expressed and confirmed by the anti-Nef antibody through western blotting. Data analyses showed that the protein possessed no considerable toxicity effect and has improved the IMT-P8 penetration rate in comparison to a control sample. Moreover, the antigen immunogenicity of the protein induced specific humoral response in mice.It was concluded that IMT-P8-Nef-MPER-V3 fusion protein has a high penetrance rate in mammalian cell line and low toxicity, thus it can be potentially considered as a vaccine against HIV-1.

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