Project description:Pseudomonas extremaustralis an Antarctic bacterium was grown at low oxygen conditions and exposed to oxidative stress during 1 hour. Experimental and control samples were analyzed by RNA-seq experiments.
Project description:sRNA40 (a novel sRNA) was pulse-expressed and its effects studied by transcriptomic profiling in P. extremaustralis, an Antarctic bacterium. For this approach we built a plasmid-based system, where sRNA40 was cloned under the control of the heterologous araBp promoter, inducible by arabinose (pBAD18-sRNA40). P. extremaustralis carrying the empty plasmid pBAD (pBAD18) was used as a control. Both strains were grown at low oxygen conditions for 24 hours. Cells were harvested after 10 min of induction with 0.1% arabinose. RNA was isolated using the Trizol method. RNA quality was analyzed on the Agilent Bioanalyzer and rRNA depletion was performed using the RiboZERO (Illumina). Samples were validated using an Agilent 2100 Bioanalyzer (Agilent Technologies). Libraries were prepared with TruSeq RNA Library Prep Kit v2 (Illumina) and sequenced with the Illumina NextSeq 550 platform with a single-end protocol.
Project description:Pseudomonas extremaustralis, an Antarctic bacterium, was grown at low oxygen conditions for 24h and then exposed to S-Nitrosoglutathione (GSNO)100 µM for 1 h. RNA from treated and control samples was isolated using the Trizol method. RNA quality was analyzed on the Agilent Bioanalyzer and rRNA depletion was performed using the RiboZERO kit (Illumina). Samples were validated using an Agilent 2100 Bioanalyzer (Agilent Technologies). Libraries were prepared with TruSeq RNA Library Prep Kit v2 (Illumina) and sequenced with the Illumina NextSeq 550 platform with a single-end protocol.
