Transcriptomic profiling of a heme-deficient mutant (hemA) of Escherichia coli after addition of CORM-3 and inactive CORM-3 (iCORM-3).

ABSTRACT: Wild type and hemA strains of Escherichia coli MG1655 were grown under anaerobiosis in batch culture in custom-made, stirred (200 rpm) 250 ml mini-fermenter vessels at 37 °C during continuous sparging with 100 % nitrogen gas. Cells were grown in defined medium (pH 7.0) containing 0.5 % glucose as the sole and limiting source of energy and carbon. The medium was supplemented with 0.1 % casamino acids and 5 % LB to support the growth of the hemA mutant. For maintenance of the hemA mutation, 50 mg/ml kanamycin were added to cultures of the heme-deficient mutant only. CORM-3 (final concentration of 100 uM) was added to wild type and mutant cultures at an OD600 of 0.2. iCORM-3 (final concentration of 100 uM) was added to only mutant cultures at an OD600 of 0.2. Samples were taken immediately prior to the addition of CORM-3 or iCORM-3 and at consecutive time-points thereafter (10, 20, 40, 60 and 120 min) for microarray analysis. Cells were harvested directly into phenol:ethanol to stabilise RNA and total RNA was purified using Qiagen’s RNeasy Mini Kit, according to the manufacturer's instructions. Time-course experiments with samples taken immediately prior to or 10, 20, 40, 60 and 120 min after addition of 100 uM CORM-3, or 100 uM iCORM-3 for the hemA mutant only, under 100 % anaerobiosis; 2 independent biological experiments were performed for each condition with 2 technical (dye swap) repeats per biological experiment.

ORGANISM(S): Escherichia coli

SUBMITTER: Robert Poole 

PROVIDER: E-GEOD-55097 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress