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PAHM4 vs PAO1 37C LB

ABSTRACT: Pseudomonas aeruginosa strains PAHM4 and PAO1 were grown at 37C on LB and RNA was hybridized on the Affymetrix P. aeruginosa chip to compare transcript differences from a BQ isolate to a well characterized wound isolate.

ORGANISM(S): Pseudomonas aeruginosa PAO1 Pseudomonas aeruginosa PAHM4 Pseudomonas aeruginosa

PROVIDER: GSE40461 | GEO | 2012/10/01

SECONDARY ACCESSION(S): PRJNA174088

REPOSITORIES: GEO

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PAHM4 vs PAO1 37C LB

Project description:Pseudomonas aeruginosa strains PAHM4 and PAO1 were grown at 37C on LB and RNA was hybridized on the Affymetrix P. aeruginosa chip to compare transcript differences from a BQ isolate to a well characterized wound isolate. Strains PAHM4 and PAO1 were grown in triplicate. Total RNA from each sample was pooled and amplified then assayed in triplicate resulting in 6 total samples. PAO1 is the control strain while PAHM4 is the experimental strain.

2012-10-01 | E-GEOD-40461 | biostudies-arrayexpress

PAHM4 vs PAO1 37C LB

Project description:PAHM4 vs PAO1 37C LB

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PAO1 gene expression at 22°C and 37°C in Lysogeny broth

Project description:Pseudomonas aeruginosa strain PAO1 was grown at 22°C and 37°C in Lysogeny broth (LB) and RNA was hybridized on the Affymetrix P. aeruginosa chip.

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PAO1-psrA::Tn expression compared to PAO1 in LB

Project description:b-Oxidative enzymes for fatty acid degradation (Fad) of long-chain fatty acid (LCFA), a component of lung surfactant phosphatidylcholine, are induced in vivo during lung infection in cystic fibrosis patients, which could contribute to nutrient acquisition and pathogenesis of Pseudomonas aeruginosa. In addition, fatty acid biosynthesis (Fab) is essential for the syntheses of two virulence controlling acylated-homoserine-lactone molecules in this organism. We mapped the promoter regions of the fadBA5-operon (PA3014 and PA3013) and a fadE homologue (PA2815) involved in Fad and the fabAB-operon involved in Fab. Focusing on the transposon mutagenesis of strain PAO1 carrying the PfadBA5-lacZ fusion, we identified a regulator for the fadBA5-operon to be PsrA (PA3006). Transcriptome analysis of the DpsrA mutant indicates its importance in regulating b-oxidative enzymes, which confirms a previous proteomic study. We further showed that induction of the fadBA-operon responds to LCFA signals, and this induction requires the presence of PsrA, suggesting that PsrA binds to LCFA to derepress fadBA5. Electrophoresis mobility shift assay indicate specific binding of PsrA to the fadBA5-promoter region. This binding is disrupted by specific LCFA (C18:1D9, C16:0, and to a lesser extent C14:0), but not by the first intermediate of b-oxidation, acyl-CoA. We proposed that PsrA is a Fad-regulator that binds and responds to LCFA signals in Pseudomonas aeruginosa. Keywords: Pseudomonas aeruginosa, wild-type PAO1 compared to PAO1-psrA::Tn

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Grad-seq in Pseudomonas aeruginosa PAO1

Project description:Protein complexome analysis of Pseudomonas aeruginosa PAO1 at OD 0.3 in LB media through 10-40% glycerol gradient fractionation in 21 samples.

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Transcription profiling of Pseudomonas aeruginosa PAO1-psrA::Tn vs. PAO1 in LB

Project description:b-Oxidative enzymes for fatty acid degradation (Fad) of long-chain fatty acid (LCFA), a component of lung surfactant phosphatidylcholine, are induced in vivo during lung infection in cystic fibrosis patients, which could contribute to nutrient acquisition and pathogenesis of Pseudomonas aeruginosa. In addition, fatty acid biosynthesis (Fab) is essential for the syntheses of two virulence controlling acylated-homoserine-lactone molecules in this organism. We mapped the promoter regions of the fadBA5-operon (PA3014 and PA3013) and a fadE homologue (PA2815) involved in Fad and the fabAB-operon involved in Fab. Focusing on the transposon mutagenesis of strain PAO1 carrying the PfadBA5-lacZ fusion, we identified a regulator for the fadBA5-operon to be PsrA (PA3006). Transcriptome analysis of the DpsrA mutant indicates its importance in regulating b-oxidative enzymes, which confirms a previous proteomic study. We further showed that induction of the fadBA-operon responds to LCFA signals, and this induction requires the presence of PsrA, suggesting that PsrA binds to LCFA to derepress fadBA5. Electrophoresis mobility shift assay indicate specific binding of PsrA to the fadBA5-promoter region. This binding is disrupted by specific LCFA (C18:1D9, C16:0, and to a lesser extent C14:0), but not by the first intermediate of b-oxidation, acyl-CoA. We proposed that PsrA is a Fad-regulator that binds and responds to LCFA signals in Pseudomonas aeruginosa. Experiment Overall Design: PAO1 and PAO1-psrA::Tn cultures grown in LB and cells were harvested at mid-log phase. Total RNA was isolated from both samples, and used for cDNA synthesis. And then, the cDNA for both samples were fragmented and labeled. The cDNA of PAO1 was used for 2 GeneChips, and PAO1-psrA::Tn cDNA was used for three GeneChips.

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PAO1 gene expression at 22M-BM-0C and 37M-BM-0C in Lysogeny broth

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