Project description:M. tuberculosis thioredoxin reductase (TrxB2) is essential for Mtb physiology and pathogenesis. To gain insight into its biological functions, we generated the TrxB2-TetON-tetO mutants, in which TrxB2 is partially depleted in the absence of atc without impairing bacterial viability. We grew TrxB2-TetON-tetO-WT in 7H9 medium with 400 ng/ml atc, until the OD reached 0.5~0.6. Then Mtb was washed with 7H9 medium 3 times and suspended in 7H9 medium with or without atc. Then samples were taken 24, 48, 72 and 120 hrs later.
Project description:M. tuberculosis thioredoxin reductase (TrxB2) is essential for Mtb physiology and pathogenesis. To gain insight into its biological functions, we generated the TrxB2-TetON-tetO mutants, in which TrxB2 is partially depleted in the absence of atc without impairing bacterial viability.
Project description:(i) RNA-seq of M. tuberculosis H37Rv cultures grown in the presence of BVL3572S for 6 h compared to untreated controls. (ii) Genome-wide M. tuberculosis H37Rv CRISPRi libraries were grown in the presence of ATc for 5 days prior to drug exposure. Cultures were treated with sub-MIC BVL3572S concentrations without amino acid or in the presence of L-His, L-Ala, or both. After 14 days outgrowth, genomic DNA were harvested from cultures for which BVL3572S concentrations were partially growth inhibitory and from untreated controls. Abundance of sgRNA was analyzed by deep sequencing. (iii) Genome-wide M. tuberculosis H37Rv transposon mutant librairies were treated with BVL3572S or untreated for 15 days followed by a 7-day antibiotic-free outgrowth. Genomic DNA were extracted, indexed, and sequenced.
Project description:Transcriptional profiling of gyr(-) strain of Mycobacterium tuberculosis H37Ra comparing 20ng/ml ATc treated cells with ATc-untreated cells after 4 days of treatment with shaking at 200rpm at 37°C.
Project description:Transcriptional profiling of Mycobacterium tuberculosis H37Rv::pTetInt-dcas9 comparing 100ng/ml ATc treated cells with Atc-untreated cells after 7 days of treatment with shaking at 200rpm at 37°C.
Project description:Transcriptional profiling of Mycobacterium tuberculosis H37Rv::pTetInt-dcas9 comparing 100ng/ml ATc treated cells with Atc-untreated cells after 4 days of treatment with shaking at 200rpm at 37°C.
