Project description:Objectives: determination of transcription start sites in Vibrio harveyi genome and discovery of new transcripts Methods: we performed differential seqencing of total RNA isolated from o.n. control Vibrio harveyi cultures. Sample treatment with Terminator EXonuclease (TEX) allowed differenciation of primary and secondary transcripts, helping in the definition of transcription start sites (TSS) Results: by data-mining RNA-seq data and performing some Northern Blot experiments we were able to detect new putative small-RNAs, along with these results, a more deep analisys of our RNA-seq data will give futher insight into genetic organization of Vibrio harveyi genome to help in its investigation Overall design: Total RNA was isolated from control o.n. cultures of Vibrio harveyi cells, strain ATCC 14126, by triplicate. RNA samples were differentialy treated with TEX and sequenced. Reads were aligned to Vibrio harveyi strain ATCC 43516 to detect TSS contributor: Vertis Biotechnologie AG
Project description:LuxR is the master quorum sensing regulator in Vibrio harveyi. This transcription factor controls the expression of over 600 genes to coordinate group behaviors. In order to understand how LuxR regulates transcription we identified a co-regulator, integration host factor (IHF). In this study we performed RNA-seq to show that the LuxR and IHF regulons overlap significantly. Overall design: Wild-type, ΔluxR, ΔihfA ΔihfB strains were grown at 30 °C to OD600 = 1.0. RNA was extracted and prepared from three biological replicates of each strain. cDNA libraries were created and sequenced for differential gene expression analysis.