M. smegmatis response to deletion of Crp1 and overexpression of Crp2
Ontology highlight
ABSTRACT: Transcriptional profiling of M. smegmatis and Dcrp1 grown in flasks on Hartmans de Bont medium supplemented with 10 mM glycerol. Transcriptional profiling of M.smegmatis HLA102 harboring tetracycline inducible vector pMind containing the Crp2 and M. smegmatis harboring pMind grown in flasks on Hartmans de Bont medium supplemented with 10 mM glycerol Comparing transcriptional response of wild-type compared with the deletion of crp1 gene (Msmeg_0539). Biological replicates: 4 independently grown and harvested. One replicate per array. Comparing transcriptional response of strain HLA102 to conditional expression of Crp2 (Msmeg_6189) compared with the wild-type with empty vector pMind. Biological replicates: 4 independently grown and harvested. One replicate per array.
Project description:Transcriptional profiling of M. smegmatis MB100pruR vs. MB100 (isogenic parent) grown in serumvials on Hartmans de Bont medium supplemented with glycerol and proline Two-condition experiment. Biological replicates: 4 independently grown and harvested. One replicate per array.
Project description:To identify the AmtR regulon of Mycobacterium smegmatis, we created a markerless deletion of the amtR gene in the background of strain M. smegmatis mc2155 (wild-type) and compared the transcription profile of both strains grown in batch culture under aerobic conditions on Hartmans de Bont medium supplemented with glycerol (carbon source) and lysine (sole nitrogen source) using microarray. Cells were harvested in early exponential growth stage.
Project description:Transcriptional profiling of M. smegmatis JR121 expressing VapC and VapBC grown in flasks on Hartmans de Bont medium supplemented with 0.2% glycerol Comparing transcriptional response of strain JR121 to conditional expression of VapC toxin compared with the expression of VapBC complex. Biological replicates: 4 independently grown and harvested. One replicate per array.
Project description:Transcriptional profiling of M. smegmatis mc2155 grown on Hartmans de Bont (HdB) supplemented with 0.2% glycerol and 0.05% Tween80 challenged with 2 μg/ml Bedaquiline and Dimethyl sulfoxide (DMSO) Two-condition experiment. Biological replicates: 5 independently grown and harvested. One replicate per array.
Project description:DosR is the regulator of a two-component system in mycobacteria that senses oxygen concentration and the redox state of the cells. To determine the DosR regulon, the transcriptomes of M. smegmatis mc2155 and the M-NM-^TdosR cells were compared by microarray at 10 hours following induction of oxygen-limitation in serum vials. 4 biological replicates incl. 2 dye swaps. 3 technical replicates per array
Project description:Comparing transciptional profile of a M. smegmatis mc2155 ÎsigF strain and wild type (control) in in vitro culture at two time points (exponential phase and stationary phase) for identification of SigF-dependent genes. Analysis of 2 time points: exponential and stationary phase. 4 biological replicates comparing expression of wild type (control) and ÎsigF strain incl. dye swaps for each time point. 3 technical replicates per array
Project description:Transcriptional profiling of M. smegmatis grown at 69h doubling time compared to 4.6h doubling time both at 50% air saturation. Transcriptional profiling of M. smegmatis grown at 69h doubling time with 50%, 2.5% and 0.6% air saturation. Two-condition experiment. Biological replicates: 5 independently grown and harvested. One replicate per array.
Project description:Mycobacterium smegmatis mc2 155 wild type strain was grown in triplicate in modified Hartmans-de Bont minimal medium (HdB) at 37C and vigorous agitation. Cultures were exposed to 1 mM NaOCl at an optical density at 500 nm (OD500) of 0.4 0.45 in the early log phase. M. smegmatis mc2 155 cells of 3 replicate experiments were harvested before and 30 min after exposure to 1 mM NaOCl and disrupted in 3 mM EDTA/ 200 mM NaCl lysis buffer with a Precellys24 Ribolyzer. RNA isolation was performed using phenol-chloroform-isoamylalcohol approach. After precipitation with 3 M sodium acetate and isopropanol, the total RNA was washed with cold ethanol and the pellet was solved in sterile water. Initially RNA quality was checked by Trinean Xpose (Gentbrugge,Belgium) and Agilent RNA Nano 6000 kit on Agilent 2100 Bioanalyzer (Agilent Technologies, Boblingen, Germany). Samples contaminated with DNA were treated with DNase (Qiagen), cleaned as described above and rechecked by Xpose and Agilent Bioanalyzer. Finally RNA was free of DNA with an RNA Integrity Number (RIN) > 9 and rRNA Ratio [23s / 16s] > 1.5. Ribo-Zero rRNA Removal Kit (Bacteria) from Illumina (San Diego, CA, USA) was used to remove the ribosomal RNA molecules from the isolated total RNA. Removal of rRNA was checked by Agilent RNA Pico 6000 kit on Agilent 2100 Bioanalyzer (Agilent Technologies, Boblingen, Germany). RNA was free of detectable rRNA. TruSeq Stranded mRNA Library Prep Kit from Illumina (San Diego, CA, USA) was used to prepare cDNA libraries. The resulting cDNAs were sequenced paired end on an Illumina HiSeq 1500 and MiSeq system (San Diego, CA, USA) using 75 bp read length.
Project description:Transcriptional profiling of M. smegmatis and Dcrp1 grown in flasks on Hartmans de Bont medium supplemented with 10 mM glycerol. Transcriptional profiling of M.smegmatis HLA102 harboring tetracycline inducible vector pMind containing the Crp2 and M. smegmatis harboring pMind grown in flasks on Hartmans de Bont medium supplemented with 10 mM glycerol
Project description:Illumina TruSeq stranded mRNA RNA-seq including Illumina ribezeo rRNA depletion of Mycobacterium smegmatis wild type and deletion mutant of the redox-sensing MarR-type repressor HypS in three bilogical replicates prior and after 30 min after exposure to 500 µM NaOCl stress revealed that hypS is autoregulated and represses transcription of the co-transcribed hypO gene which encodes a multidrug efflux pump. DNA binding activity of HypS to the 8-5-8 bp inverted repeat sequence upstream of the hypSO operon was inhibited under NaOCl stress. HypS was identified as a novel redox-sensitive repressor that contributes to mycobacterial resistance towards HOCl stress and antibiotics.