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  • Genes required for and effects of alginate overproduction induced by growth of Pseudomonas aeruginosa on Pseudomonas isolation agar supplemented with ammonium metavanadate.

Genes required for and effects of alginate overproduction induced by growth of Pseudomonas aeruginosa on Pseudomonas isolation agar supplemented with ammonium metavanadate.

Journal of bacteriology (2013-06-25)
F Heath Damron, Mariette Barbier, Elizabeth S McKenney, Michael J Schurr, Joanna B Goldberg
ZUSAMMENFASSUNG

Pseudomonas aeruginosa is an opportunistic pathogen that can adapt to changing environments and can secrete an exopolysaccharide known as alginate as a protection response, resulting in a colony morphology and phenotype referred to as mucoid. However, how P. aeruginosa senses its environment and activates alginate overproduction is not fully understood. Previously, we showed that Pseudomonas isolation agar supplemented with ammonium metavanadate (PIAAMV) induces P. aeruginosa to overproduce alginate. Vanadate is a phosphate mimic and causes protein misfolding by disruption of disulfide bonds. Here we used PIAAMV to characterize the pathways involved in inducible alginate production and tested the global effects of P. aeruginosa growth on PIAAMV by a mutant library screen, by transcriptomics, and in a murine acute virulence model. The PA14 nonredundant mutant library was screened on PIAAMV to identify new genes that are required for the inducible alginate stress response. A functionally diverse set of genes encoding products involved in cell envelope biogenesis, peptidoglycan remodeling, uptake of phosphate and iron, phenazine biosynthesis, and other processes were identified as positive regulators of the mucoid phenotype on PIAAMV. Transcriptome analysis of P. aeruginosa cultures growing in the presence of vanadate showed differential expression of genes involved in virulence, envelope biogenesis, and cell stress pathways. In this study, it was observed that growth on PIAAMV attenuates P. aeruginosa in a mouse pneumonia model. Induction of alginate overproduction occurs as a stress response to protect P. aeruginosa, but it may be possible to modulate and inhibit these pathways based on the new genes identified in this study.

MATERIALIEN
Produktnummer
Marke
Produktbeschreibung

Sigma-Aldrich
Agar, microbiology tested, suitable for plant cell culture, suitable for cell culture, powder
Sigma-Aldrich
Ammoniummetavanadat, ACS reagent, ≥99.0%
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Natriumalginat, powder
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Natriumalginat
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Agar, suitable for plant cell culture, powder
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Natriumalginat aus Braunalgen, low viscosity
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Natriumalginat aus Braunalgen, Medium viscosity
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Ammoniummetavanadat, puriss. p.a., ACS reagent, ≥99.0% (RT)
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Natriumalginat aus Braunalgen, BioReagent, suitable for immobilization of micro-organisms
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Alginsäure aus Braunalgen, powder
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Agar, Bacteriological, microbiology tested, suitable for plant cell culture, granular
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Agar, High gel strength, suitable for plant cell culture
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Ammoniummetavanadat, 99.95% trace metals basis
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Agar, Type A, suitable for plant cell culture
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Agar, powder, suitable for microbiology
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Calciumalginat aus Braunalgen
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Agar, Type M, suitable for plant cell culture
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Bakteriologischer Agar, for molecular biology
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Agar, suitable for plant cell culture
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Noble-Agar
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Agar, ash 2.0-4.5%
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Select-Agar
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Agar, suitable for microbiology
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Agar, high purity, low ionic content, low gel strenght, suitable for microbiology