Characterisation of iunH gene knockout strain from Mycobacterium tuberculosis.

摘要: Background Tuberculosis (TB) is an infectious disease caused mainly by the bacillus Mycobacterium tuberculosis. The better understanding of important metabolic pathways from M. tuberculosis can contribute to development novel therapeutic and prophylactic strategies combat TB. Nucleoside hydrolase (MtIAGU-NH), encoded iunH gene (Rv3393), enzyme purine salvage pathway in MtIAGU-NH accepts inosine, adenosine, guanosine, uridine as substrates, which may point a pivotal role. Objectives Our aim was construct knockout strain for gene, evaluate vitro growth effect deletion non-activated activated macrophages models infection. Methods A obtained allelic replacement, using pPR27xylE plasmid. complemented constructed transformation with pNIP40::iunH. expression analysed Western blot LC-MS/MS. In evaluated Sauton's medium. Bacterial load interferon-γ RAW 264.7 cells infected compared wild-type strains. Findings LC-MS/MS validated at protein level. led delay kinetics medium during log phase, but did not affect bases nucleosides pool vitro. No significant difference bacterial observed when both strains after infection cells. Main conclusion disruption does influence cells, show that macrophage invasion virulence. results indicated target drug development.