Project description:The goal was to identify genes regulated by YvcL (EF0768) by assessing changes in the yvcL deletion mutant relative to the isogenic wild-type. Wild-type E. faecalis OG1 and the mutant lacking yvcL were cultured in Mueller-Hinton broth to exponential phase. Total RNA was purified, depleted for rRNA, and subjected to preparation of mRNA libraries.
Project description:To globally characterize the role of rnjB gene in E.faecalis, the transcriptome of ∆rnjB and OG1RF grown to exponential phase were compared using DNA microarray analysis
Project description: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).
Project description:To further investigate the homeostatic response of E. faecalis to Fe exposure, we examine the whole-genome transcriptional response of wild-type (WT) exposed to non toxic Fe excess. This experiment correspond the work titled Transcriptomic response of Enterococcus faecalis to iron excess (work in preparation)
Project description:The enterococci comprise a genus of 49 low-GC content Gram-positive commensal species within the Firmicutes phylum that are known to occupy diverse habitats, notably the gastrointestinal core microbiota of nearly every phylum, including human. Of particular clinical relevance are two rogue species of enterococci, Enterococcus faecalis and the distantly related Enterococcus faecium, standing among the nefarious multi-drug resistant and hospital-acquired pathogens. Despite increasing evidence for RNA-based regulation in the enterococci, including regulation of virulence factors, their transcriptome structure and arsenal of regulatory small sRNAs (sRNAs) are not thoroughly understood. Using dRNA-seq, we have mapped at single-nucleotide resolution the primary transcriptomes of E. faecalis V583 and E. faecium AUS0004. We identified 2517 and 2771 transcription start sites (TSS) in E. faecalis and E. faecium, respectively. Based on the identified TSS, we created a global map of s70 promoter motifs. We also revealed features of 5’ and 3’UTRs across the genomes. The transcriptome maps also predicted 150 and 128 sRNA candidates in E. faecalis and E. faecium, respectively, some of which have been identified in previous studies and many of which are new. Finally, we validated several of the predicted sRNAs by Northern Blot in biologically relevant conditions. Comprehensive TSS mapping of two representative strains will provide a valuable resource for the continued development of RNA biology in the Enterococci.
