Project description:A M. tuberculosis transposon library was used to infect WT and iNOS-/- mice. Surviving mutants were recovered from spleens, genomic DNA was extracted, and labeled probes were synthesized from transposon ends. Probes from each WT mouse were hybridized with probes from a similar iNOS-/- mouse. Two-condition experiment, Growth in WT vs. iNOS-/- mice. Biological replicates: TraSH probe made from 5 wild type and 5 iNOS-/- mice after 3 weeks of infection and from 8 wild type and 8 iNOS-/- mice after 4 weeks of infection. Technical replicates: TraSH probe was synthesized twice from each mouse and dyes were swapped. One array contains probe from one iNOS-/- and one WT mouse. Each biological replicate has two arrays, representing technical replicate dye swaps from one iNOS-/- and one WT mouse.
Project description:Mycobacterium tuberculosis transposon mutants with reduced bodipy-palmitate intake on the third day of resting bone marrow-derived macrophage infection are identified by TraSH. Discovery of genetic requirements for fatty acid assimilation by the pathogen is essential for understanding its metabolism during host infection.
Project description:We used single-cell RNA sequencing (scRNAseq) to compare cells from the lungs of Mycobacterium tuberculosis infected mice between C57BL/6J mice and a cohort of wild-derived mouse lines MANA, MANB and MANC (Jackson Laboratories: ManA/NachJ #035354; ManB/NachJ #035355).
Project description:Mycofactocin is a new class of peptide-derived redox cofactor whose structural elucidation and functional characterization have recently received enormous focus. Among the six-gene cluster, mftD mediates a probable penultimate step in mycofactocin biosynthesis. MftD is a putative lactate dehydrogenase predicted critical for Mycobacterium tuberculosis survival under oxygen-limited conditions. In this study, mftD transcripts levels were found significantly increased in M. tuberculosis cells adapted to 0.01% oxygen. mftD deletion mutant of M. tuberculosis exhibited survival deficit in in vitro hypoxia and wayne models. However, mftD functions was found dispensable for M. tuberculosis L-Lactate metabolism. Rather surprisingly, the growth fitness of mftD mutant was increased in glucose under aerobic conditions. While the cause of this in vitro phenotype remains unestablished, the levels of NAD(P)H and glucose-6-phosphate dehydrogenase activity was found decreased in ΔmftD when compared to its parental strain. Increased growth fitness did not have any major impact on bacterial cell shape and size except the formation of extracellular fibril-like structures in subpopulations of ΔmftD. Cell-surface proteins analysis showed that genetic deficiency of mycofactocin likely to predispose accumulation of cofactor-free protein aggregates resembling destabilization of the flavoproteome upon riboflavin deprivation. Nevertheless, like in vitro findings, disruption of mftD increased the fitness of M. tuberculosis in C57BL/6J mice at early phase and resulted in growth stasis in a Nos2-/- hypoxia mouse model. Collectively these results establish the relevance of MftD in M. tuberculosis growth, redox and cofactor homeostasis, and pathogenesis.
Project description:Transcription analysis of M-NM-^TpknK mutant (LIX11) versus wild type H37Rv during logarithmic and stationary phase growth. This will elucidate the genes regulated by PknK during transition from logarithmic growth to stationary phase growth. Two color Experiment,Organism: Mycobacterium tuberculosis, Genotypic Technology designed Custom Mycobacterium tuberculosis H37Rv Whole Genome 8x15k GE Microarray (AMADID-26323 and AMADID-23057
Project description:Transcriptional profiling of SirR and manganese regulated expression of genes in Mycobacterium tuberculosis strains comparing high manganese vs. low manganese in Rv (wild type Mycobacterium tuberculosis) and ST70 (mntR mutant strain of Mycobacterium tuberculosis) Two strains each with two conditions experiment, Rv (Mycobacterium tuberculosis wild type strain) high manganese vs. low manganese and ST70 (mntR mutant strain of Mycobacterium tuberculosis) high manganese vs. low manganese. Number of biological replicates is 3 for each condition for each strain.
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:Transcriptional profiling of Mycobacterium tuberculosis H37Rv strains comparing control DMSO treated strains with Lupulone treated strains. Goal was to determine the effects of Lupulone against Mycobacterium tuberculosis H37Rv strains. Two-condition experiment,control DMSO treated strains vs. Lupulone treated strains. Biological replicates: 2 control replicates, 2 Lupulone replicates.
Project description:Transcriptional profiling of Mycobacterium tuberculosis H37Rv strains comparing control DMSO treated strains with Linezolid treated strains. Goal was to determine the effects of Linezolid against Mycobacterium tuberculosis H37Rv strains. Two-condition experiment,control DMSO treated strains vs. Linezolid treated strains. Biological replicates: 2 control replicates, 2 Linezolid replicates.
Project description:This SuperSeries is composed of the following subset Series: GSE36341: mRNA degradation in Mycobacterium tuberculosis under aerobic conditions GSE36342: mRNA degradation in Mycobacterium smegmatis under aerobic conditions GSE36343: mRNA degradation in Mycobacterium tuberculosis during cold and hypoxic stress GSE36344: mRNA degradation in Mycobacterium tuberculosis with DosR ectopically induced Refer to individual Series
