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Real‐time PCR used to measure stress‐induced changes in the expression of the genes of the alginate pathway of Pseudomonas aeruginosa
Author(s) -
Edwards K.J.,
Saunders N.A.
Publication year - 2001
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1046/j.1365-2672.2001.01339.x
Subject(s) - pseudomonas aeruginosa , gene expression , gene , biology , real time polymerase chain reaction , messenger rna , microbiology and biotechnology , overproduction , heat shock protein , regulation of gene expression , regulator gene , bacteria , biochemistry , genetics
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.