Rv0500A is a transcription factor that links Mycobacterium tuberculosis environmental response and division

The purpose of this study was to identify genes that are differentially expressed upon rv0500A over-expression, to reveal the impact of Rv0500A on global transcription in different environmental conditions. Mtb strain Erdman harboring a tetracycline inducible N-terminally tagged 6x His-Rv0500A construct was grown to an OD600 of 0.6 in standing, filter-capped T-75 flasks and treated with either 200 ng/ml anhydrotetracycline (ATC) or ethanol (EtOH) as a carrier control for 2 hours. Bacteria were then subcultured to an OD600 of 0.3 in standing, filter-capped T-25 flasks in either 7H9, pH 7 media, or 7H9, pH 5.7 media supplemented with 250 mM NaCl, with continued presence of ATC or EtOH as appropriate. Exposure to the different media conditions was for 4 hours in a humidified, 37°C, 5% CO2 incubator before RNA was extracted. RNA was prepared by removing the rRNA and library prepping with a TruSeq Stranded kit (Illumina) before high-throughput sequencing with an Illumina HiSeq 2500 (High Output v4) (100 bp single end reads, one lane). Two biological samples per condition were sequenced.