Project description:Transcriptional profiling of mosquito head comparing control non-swarming mosquito with swarming mosquito. Screen the genes associated with swarming.
Project description:Four biological repeats of P.aeruginosa PA14 and four biological repeats of P.aeruginosa PA14 metR::phoA were grown in BM2 swarming media plates containing 62mM potassium phosphate buffer [pH7], 2 mM MgSO4, 10uM FeSO4, 0.4% (wt/vol) glucose, 0.1% (wt/vol) casamino acids and 0.5% agar. Microarray experiments were done using microarray slides and protocols from TIGR on samples harvested from the dendritic swarm colony growth, 2 to 3 mm of the swarm zone edge.
Project description:In this experiment the transcriptional profile of the Pseudomonas aeruginosa PA14 two-component sensor kinase PA4398 was investigated under swarming conditions using DNA microarrays. To this aim three independent cultures of the PA14 wild-type and the PA4398 mutant were grown until mid-log phase in Luria-Bertani broth following an incubation on BM2-swarm plates containing 0.1 % (wt/vol) casaminoacids and 0.5 % (wt/vol) agar for 20 h at 37 °C. Subsequent total RNA was extracted from the leading edge of dendritic swarm colonies and analyzed by microarrays.
Project description:Abstract: Pseudomonas aeruginosa is a Gram-negative opportunistic pathogen found ubiquitously in the environment. Responsible for considerable human morbidity and mortality, particularly in nosocomial infections and individuals with cystic fibrosis, P. aeruginosa can adapt to surface growth by undergoing swarming motility. In P. aeruginosa, swarming motility is a rapid multicellular movement that occurs on soft surfaces of appropriate viscosity with amino acids as a nitrogen source. Here we tested the small synthetic host defense peptide, innate defense regulator 1018, and found that it inhibited swarming motility at concentrations as low as 0.75 μg/ml, well below the MIC for planktonic cells. A screen of the PA14 transposon insertion mutant library revealed twenty-nine mutants that demonstrated partial tolerance to 1018 under swarming conditions. Two of these mutants, in the genes anr and rhlB (a regulator of anaerobic metabolism and protein involved in rhamnolipid production, respectively), were complemented to restore susceptibility to 1018. RNA-Seq of peptide-treated cells under swarming conditions revealed the dysregulation of 1,190 genes compared to the untreated swarm front, and 67% of these genes were similarly dysregulated at the untreated swarm centre. In contrast, expression of 70 Anr-regulated genes was upregulated by peptide treatment, and 45 genes showed differential or opposite regulation of expression in peptide-treated and swarm centre cells. Many transcriptional regulators required for swarming were dysregulated in peptide-treated cells, indicative of a mechanism by which 1018 may inhibit swarming motility. Overall, this study illustrates a use for peptide 1018 in inhibiting swarming surface motility, an important bacterial adaptation.
Project description:Salmonella 14028 time course on swarming agar (0.6%) plates mRNA from 3 h time point on swarming plates used as reference for all the time course arrays Keywords: other
Project description:Salmonella strain 14028, time course, on non-swarming agar plates (1.5%), array were done between reference mRNA (from swarmer cells at 3 hour) and mRNA from various time points on non-swarming plates. Keywords: other
Project description:Swarming motility by the urinary tract pathogen Proteus mirabilis has been a long-studied, but little understood phenomenon. On agar, a P. mirabilis colony grows outward in a bullseye pattern formed by consecutive waves of rapid swarming followed by consolidation into shorter cells. To examine differential gene expression in these growth phases, a microarray, constructed based on the completed genome sequence and annotation, was undertaken. RNA from 1) broth-cultured, or 2) swarming cells was extracted to assess transcription during each of these growth states.
Project description:Salmonella 14028 time course on swarming agar (0.6%) plates mRNA from 3 h time point on swarming plates used as reference for all the time course arrays