E-MTAB-1465 - Transcription profiling by array of mycelia from Streptomyces coelicolor A3(2) and its rok&B& null mutant grown on mannitol and isolated from different time-points in the reproductive cycle
Released on 14 January 2014, last updated on 3 May 2014
Streptomyces coelicolor A3(2)
For transcript analysis, total RNA was isolated as described (Noens et al, 2007) from surface-grown mycelia of S. coelicolor M145 and its rok7B7 null mutant, cultivated on MM agar plates with mannitol (1% w/v) as the sole carbon source (plates were covered with cellophane discs). Two independent biological replicate experiments were performed. Samples were taken at 14h (early vegetative growth), 24 h (vegetative growth), 30 h (early aerial growth), 36 h (aerial growth), 42 h (early sporulation) and 54 h (sporulation). Total RNA was purified using the Kirby-mix protocol (Kieser et al, 2000). DNaseI treatment was used to fully remove any traces of DNA. Before use, the RNA samples were checked for integrity on an Agilent 2100 Bioanalyzer (Agilent Technologies). cDNA preparation and labeling was performed as previously described (Bucca et al, 1997, Bucca et al, 2009). The filtered data sets were analyzed using rank products analysis (Breitling et al, 2004) via the Rank-ProdIt tool (Laing and Smith, 2010). Differentially expressed genes were identified as demonstrating a > 2 fold change in expression between the parental strain M145 (wild-type) and the rok7B7 mutant.
transcription profiling by array, co-expression, reference, replicate, strain or line, time series