Real-time PCR used to measure stress-induced changes in the expression of the genes of the alginate pathway of Pseudomonas aeruginosa

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Abstract
Aims: To measure the concentration of mRNAs transcribed from four genes involved in alginate production using real‐time PCR. Methods and Results: The mRNA concentrations in cells grown in normal and stress conditions were compared. A difference in the expression of algD, the key gene leading to overproduction of alginate, was detected between alginate‐producing and non‐alginate‐producing strains grown under normal conditions. After growth on 3% ethanol (known to stimulate alginate production), but not after heat‐shock, an increase in algD mRNA levels and a corresponding decrease in mucB (a regulatory gene) mRNA levels were detected in all strains. Conclusions: The quantitative results suggest that the mucB gene may have a role in recognition of stress conditions, and that having a disrupted mucA gene does not always result in a mucoid phenotype. Significance and Impact of the Study: Real‐time PCR can be used to quantify mRNA and is a convenient method of analysing bacterial gene expression.