Efficient expression of Gag protein by recombinant Modified Vaccinia Ankara Virus (MVA) with HIV-1Indian Subtype C gagprotease gene segments

摘要

The AIDS epidemic in the developing world particularly in India represents a major threat where the infections are due to non-B clades of HIV-1. Most human immunodeficiency virus (HIV) vaccines currently under development are based on clade B strains of HIV-1. Since in India clade C is the predominant strain of HIV-1, it is imperative to develop an effective vaccine candidate from the locally circulating HIV strains. We describe here the development of recombinant Modified Vaccinia Ankara (MVA) expressing gag protein of HIV-1 Indian subtype C. Plasmid transfer vector, pSC 65, was used to transfer gag-protease gene segment of HIV-1 subtype C strain 49587 to MVA and gene segment was placed under the control of synthetic early late promoter (PE/L). The recombinant MVA was selected by BrdU/X-gal selection. The recombinant MVA was found to express gag protein in BHK-21 cells and the expression was evaluated by p24 antigen capture ELISA, immunoblotting, immunoflourescence and formation of virus-like-particles (VLPs) in infected cells by electron microscopy. The construct showed stable and high expression of HIV-1 gag gene in eukaryotic cells.