CroR-regulated genes in Enterococcus faecalis OG1

The goal was to identify genes that are regulated by cell wall stress (bacitracin treatment) in wild-type cells but not in mutant cells lacking CroR, thereby revealing genes of the CroR regulon. Cells of a mutant lacking CroR or its otherwise isogenic wild-type E. faecalis OG1 were cultured in Mueller-Hinton broth and treated (or not) with bacitracin for 15 min. Total RNA was purified, depleted for rRNA, and subjected to preparation of TruSeq stranded mRNA libraries before sequencing on a NextSeq 550 (76-bp single read run). Overall design: Four replicates per condition (wild-type untreated, wild-type treated, CroR mutant untreated, CroR mutant treated). Cells cultured at 37 C until exponential phase, then treated or not with bacitracin at 128 ug/ml for 15 min before RNA extraction.