Reporter genes and fluorescent probes for studying the colonisation of biofilms in a drinking water supply line by enteric bacteria.
Biofilms containing diverse microflora were developed on bitumen-painted steel and glass tiles suspended in a chemostat model of a water distribution system. Escherichia coli, taken from a naturally occurring biofilm, was transformed with a plasmid containing the anaerobically induced nirB promoter fused to the lacZ reporter gene. The resulting transformant, PRB1, was introduced into the chemostat. After 7 and 13 days, an E. coli strain with an anaerobically induced Lac+ phenotype was present in the biofilm. Development of an episcopic differential interference contrast technique combined with UV fluorescence microscopy enabled the simultaneous visualization of E. coli in the biofilm using a fluorescent probe to detect expression of the gusA reporter gene and a lacZ fluorescent probe to monitor anaerobic expression of beta-galactosidase from pnirB.
Robinson
P J
PJ
School of Biochemistry, University of Birmingham, UK.
Walker
J T
JT
Keevil
C W
CW
Cole
J
J
D018441:Biofilms; D015415:Biomarkers; D004926:Escherichia coli / Q000235:genetics / Q000302:isolation & purification*; D005456:Fluorescent Dyes; D017930:Genes, Reporter; D014881:Water Supply
D015415:Biomarkers; D005456:Fluorescent Dyes