title: Real-time PCR used to measure stress-induced changes in the expression of the genes of the alginate pathway of Pseudomonas aeruginosa.
abstract: 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.
authors.affiliation: Molecular Biology Unit, Virus Reference Division, Central Public Health Laboratory, 61 Colindale Avenue, London NW9 5HT, UK.
authors.forename: K J
authors.initials: KJ
authors.lastname: Edwards
authors.forename: N A
authors.initials: NA
authors.lastname: Saunders
mesh_terms: 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
chemical_list: 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