Expression, purification, and functional characterization of soluble recombinant full-length simian immunodeficiency virus (SIV) Pr55Gag

• DOI: 10.1016/j.heliyon.2023.e12892
• Publication Date: 2023-01-10T01:48:49Z
• Authors: Vineeta N. Pillai; Lizna Mohamed Ali; Suresha G. Prabhu; Anjana Krishnan; Saeed Tariq; Farah Mustafa; Tahir A. Rizvi
• Keywords: Simian immunodeficiency virus; Group-specific antigen

The simian immunodeficiency virus (SIV) precursor polypeptide Pr55Gag drives viral assembly and facilitates specific recognition packaging of the SIV genomic RNA (gRNA) into particles. While several studies have tried to elucidate role by expressing its different components independently, using full-length not been conducted, primarily due unavailability purified biologically active Pr55Gag. We successfully expressed soluble, with His6-tag in bacteria it affinity gel filtration chromatography. In process, we identified within Gag, a second in-frame start codon downstream putative Shine-Dalgarno-like sequence resulting an additional truncated form Gag. Synonymously mutating this allowed expression Gag native form. assembled virus-like particles (VLPs) vitro presence nucleic acids, revealing biological functionality. vivo experiments also confirmed formation functional VLPs, quantitative reverse transcriptase PCR demonstrated efficient gRNA these VLPs. methodology employed ensured availability >95% pure, active, which should facilitate future understand protein structure RNA-protein interactions involved during packaging.