Transcriptome, inflammatory and oxidative properties of Clostridium butyricum CB1002 clinical strain and its isogenic mutant CB1002 Δhbd
Transcriptome, inflammatory and oxidative properties of Clostridium butyricum CB1002 clinical strain and its isogenic mutant CB1002 Δhbd
samples were subcultured on columbia cystein agar at 37°C for 48 hours, and then cultured in 10 mL BHI broth firstly and 20 mL BHI broth secondly overnight at 37°C prior to isolation. All cultures have been perfomed in an anaerobic chamber (Coy Laboratory Products Inc, Grass Lake, MI, USA) (v/v: 90% N2, 5% H2, 5% CO2). Clostridium butyricum cultures were incubated until it reached an optical density (OD600) of 1.1 for isolation at mid-exponential phase. Total bacterial RNA was then extracted and purified using ZYMO Direct-zol™ RNA Miniprep Plus R2070 extraction kit and treated with DNAse I according to manufacturer's instructions. A total of 100 ng RNA sample was used to construct each RNAseq library using Illumina stranded Total RNA Ligation Ribo0+ kit and subjected to ribosomal RNA (rRNA) depletion following manufacturer's instructions