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Analysis of antibiotic resistance gene expression in Pseudomonas aeruginosa by quantitative real-time-PCR
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Analysis of antibiotic resistance gene expression in Pseudomonas aeruginosa by quantitative real-time-PCR
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Analysis of antibiotic resistance gene expression in Pseudomonas aeruginosa by quantitative real-time-PCR
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Journal Article
Analysis of antibiotic resistance gene expression in Pseudomonas aeruginosa by quantitative real-time-PCR
2006
Overview
Abstract
In Pseudomonas aeruginosa many of the clinically relevant resistance mechanisms result from changes in gene expression as exemplified by the Mex drug efflux pumps, the AmpC β-lactamase and the carbapenem-specific porin OprD. We used quantitative real-time-PCR to analyze the expression of these genes in susceptible and antibiotic-resistant laboratory and clinical strains. In nalB mutants, which overexpress OprM, we observed a four- to eightfold increase in the expression of mexA, mexB, and oprM genes. MexX and mexY genes were induced eight to 12 times in the presence of 2 mg L−1 tetracycline. The mexC/oprJ and mexE/oprN gene expression levels were increased 30- to 250-fold and 100- to 760-fold in nfxB and nfxC mutants, respectively. We further found that in defined laboratory strains expression levels of ampC and oprD genes paralleled β-lactamase activity and OprD protein levels, respectively. Our data support the use of quantitative real-time-PCR chain reaction for the analysis of the antimicrobial resistance gene expression in P. aeruginosa.
Publisher
Blackwell Publishing Ltd,Blackwell Science Ltd,Blackwell,Oxford University Press
Subject
Anti-Bacterial Agents - pharmacology
/ Bacterial Proteins - genetics
/ Bacterial Proteins - metabolism
/ beta-Lactamases - metabolism
/ Biological and medical sciences
/ Drug Resistance, Bacterial - genetics
/ Efflux
/ Fundamental and applied biological sciences. Psychology
/ Gene Expression Regulation, Bacterial
/ Genes
/ Genetics
/ Humans
/ Pseudomonas aeruginosa - drug effects
/ Pseudomonas aeruginosa - genetics
/ Pseudomonas aeruginosa - metabolism
