Project description:Transcription analysis of ΔnarL mutant (LIX75) versus wild type H37Rv during aerobic growth in absence or presence of 5 mM Nitrate, and during hypoxic growth. This will elucidate the regulatory role of NarL response regulator duirng aerobic growth in absence of presence of exogenous nitrate supplemen and during hypoxic growth conditions and help identify genes that require NarL for expression under the three experimental conditions.
Project description:Transcription analysis of M-NM-^TnarL mutant (LIX75) versus wild type H37Rv during aerobic growth in absence or presence of 5 mM Nitrate, and during hypoxic growth. This will elucidate the regulatory role of NarL response regulator duirng aerobic growth in absence of presence of exogenous nitrate supplemen and during hypoxic growth conditions and help identify genes that require NarL for expression under the three experimental conditions. Two color Experiment,Organism: Mycobacterium tuberculosis, Genotypic Technology designed Custom Mycobacterium tuberculosis H37Rv Whole Genome 8x15k GE Microarray (AMADIDAMADID-23057)
Project description:Expression profile of Mycobacterium tuberculosis H37Rv biofilm as induced by DTT (Reduced) 6mM DTT reduced at 6 mM concentration was added to log phase culture of Mtb H37Rv. After 29 hours RNA was isolated and hybridization was done on microarrays
Project description:Identification of glnR-dependent regulation of nitrate assimilation Keywords: genetic modification Mycobacterium tuberculosis H37Rv wild type and glnR mutant strain were incubated in minimal medium supplemented with 5 mM NO3 for 18 and 48 hours. RNA was extracted at indicated time points and microarray analysis was performed. Experiments were repeated once. As we wanted to focus on genes that were regulated steadily over time, data obtained from time points 18 hours and 48 hours were grouped. Thus for wild type and mutant, four independent data sets were obtained each, and compared using t-test statistics.
Project description:Transcriptional profiling of Mycobacterium tuberculosis H37Rv and Mycobacterium bovis Ravenel before (i.e., log-phase control) and at 24h and 96h after nutrient starvation. The transcriptional profile of the two strains were compared at log phase, and after 24h and 96h of starvation. In addition, the response of each strain to starvation was examined after 24h and 96h of starvation.
Project description:We used three M. tuberculosis strains (WT H37Rv, an RV2752c deletion mutation, and the mutant complemented with an Rv2752c overexpression construct), extracted RNA from log phase cultures, and made RNAseq expression libraries as well as 5'-end-directed libraries.
Project description:New chemotherapeutics are urgently required to control the tuberculosis pandemic fueled by the emergence of multidrug- and extensively-drug-resistant Mycobacterium tuberculosis strains and the bacterium`s catastrophic alliance with HIV. Here we describe a novel trehalose-to-α-glucan pathway in M. tuberculosis comprising four enzymatic steps mediated by TreS, Pep2, GlgB, and GlgE, identified as an essential maltosyltransferase capable of utilizing maltose 1-phosphate. Using traditional and chemical reverse genetics, we show that GlgE inactivation causes rapid death of M. tuberculosis in vitro and in mice, through self-poisoning by maltose 1-phosphate accumulation driven by a self-amplifying feedback loop promoting pleiotropic phosphosugar-induced stress responses. Moreover, this α-glucan pathway exhibited a synthetic lethal interaction with the glucosyltransferase Rv3032 involved in biosynthesis of specialized α-glucan derivatives. The unique combination of gene essentiality within a synthetic lethal pathway validates GlgE as a new class of drug targets, revealing novel synergistic mechanisms to induce death in M. tuberculosis. Transcriptional profiling was performed to characterize the lethality induced by maltose 1-phosphate accumulation. Triplicate 10 mL cultures of the conditional lethal Mtb mutant strain H37Rv Delta treS Delta glgE (pMV361::treS) and of the vector control strain H37Rv Delta treS Delta glgE (pMV361) were grown in liquid culture to log-phase in the presence of 5 mM validamycin A (VA) to suppress M1P formation. Subsequently, cells were washed to remove the inhibitor; after 48 h of starvation for VA cultures were harvested. Keywords: tuberculosis, trehalose, compound treatment design, genetic modification design, and stimulus or stress design
