Real-time PCR used to measure stress-induced changes in the expression of the genes of the alginate pathway of Pseudomonas aeruginosa.
OBJECTIVE
To measure the concentration of mRNAs transcribed from four genes involved in alginate production using real-time PCR.


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.


CONCLUSIONS
Real-time PCR can be used to quantify mRNA and is a convenient method of analysing bacterial gene expression.
Edwards
K J
KJ
Molecular Biology Unit, Virus Reference Division, Central Public Health Laboratory, 61 Colindale Avenue, London NW9 5HT, UK.
Saunders
N A
NA
D000464:Alginates / Q000378:metabolism*; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics; D001483:Base Sequence; D002237:Carbohydrate Dehydrogenases / Q000235:genetics / Q000378:metabolism; D004252:DNA Mutational Analysis; D000431:Ethanol / Q000494:pharmacology; D017353:Gene Deletion; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects; D020723:Glucuronic Acid; D006603:Hexuronic Acids; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods; D011550:Pseudomonas aeruginosa / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000187:drug effects / Q000378:metabolism; D013696:Temperature
D000464:Alginates; D001426:Bacterial Proteins; D006603:Hexuronic Acids; C063757:MucA protein, Bacteria; D012333:RNA, Messenger; D000431:Ethanol; D020723:Glucuronic Acid; D002237:Carbohydrate Dehydrogenases; C020562:GDPmannose dehydrogenase