The impact of cobB1 or cobB2 gene deletions on global transcript levels in Streptomyces venezuelae
The impact of cobB1 or cobB2 gene deletions on global transcript levels in Streptomyces venezuelae
RNA was isolated from Streptomyces venezeulae JD01 (cobbB1 deletion) or JD07 (cobB2 deletion) strains cultivated in 5 mL MYM liquid medium for 10 or 20 hours (each time point in duplicate). Mycelia were collected by centrifugation, frozen, and stored at -70Â°C for subsequent RNA isolation. RNA from mycelia was isolated using the GeneJET RNA isolation kit (Thermo Fisher Scientific, US). The lysozyme concentration in the suspension buffer was increased up to 10 mg/mL to improve the yield of mycelium lysis. The isolated RNA was digested with TURBO DNase I (Invitrogen, United States) and subsequently, purified and concentrated using a GeneJET RNA Cleanup kit (Thermo Fischer Scientific, US) RNA libraries were prepared for sequencing using standard Illumina protocols (NEBNext Ultra II Directional RNA Library Prep Kit). rRNA was removed using Ribo-Zero Ribosomal RNA Removal Kit (Illumina)