RNA-Seq of Streptomyces venezuelae (pxd): 48h after ethanol shock
Streptomyces venezuelae NRRL B-65442 and deletion mutants pepX, RS15950, RS22785 and RS24280 were each grown in MYM medium, containing 2.1 g/L MOPS (Sigma). Ethanol shock was performed after 7 h of cultivation by adding 1.5 mL (6% v/v; only wt and pepX) of absolute ethanol or 1.5 mL distilled sterile water (control) to the cultures. The fermentations were stopped after 18, 24 and 48 h (only at 24h for RS15950, RS22785 and RS24280). All the fermentations have been performed in triplets. 1.5 mL samples were immediately taken from each culture and transferred in sterile RNAse-free tubes containing 3 mL RNAprotect Bacteria Reagent (QIAGEN, Germany). The samples were vortexed for 10 sec, rested on table for 5 min, transferred into 2.5 mL sterile RNAse-free Eppendorf tubes and centrifuged for 12 min at 4000 rpm, 4 C. Supernatants were removed and cells frozen immediately at -80 C. One tube with the cells per sample was taken for RNAseq analysis.