E-MTAB-4833 - Deciphering the transcriptional response mediated by the redox-sensing system HbpS-SenS-SenR from streptomycetes
Last updated on 20 June 2016, released on 30 June 2016
Comparative analyses of the transcriptomes of the Streptomyces coelicolor A3(2) wild-type and the generated hbpSc-senSc-senRc (hsr) mutant under native and oxidative-stressing conditions allowed to identify differentially expressed genes, whose products may enhance the anti-oxidative defense of the bacterium. To obtain well-grown mycelia, spores of S. coelicolor (WT and hsr mutant) were inoculated in 10 ml R2 medium and grown as standing culture at 30C for 16 h and afterwards on a rotary shaker for 16 h after the addition of 90 ml R2 medium. The cultures were washed four times in minimal medium without supplement. The mycelia were suspended in 50 ml R2 medium and divided in two 25 ml-portions, one of which contained H2O2 (0.15 mM). Cultivation was continued at 30 C on a rotary shaker for two hours. Mycelia were harvested by centrifugation and the mycelia pellets were store at -80 C. Total RNA was isolated from two biological replicates. cDNA libraries of RNA from of S. coelicolor (WT and hsr mutant) were constructed using the TruSeq Stranded mRNA Library Prep Kit (Illumina,San Diego, CA, USA), and subsequently sequenced paired-end on an Illumina MiSeq system (San Diego, CA, USA) using 75 bp read length.
RNA-seq of coding RNA, genetic modification design, genotype design, stimulus or stress design
Anika Winkler, Tobias Busche <firstname.lastname@example.org>