Open AccessReporter genes and fluorescent probes for studying the colonisation of biofilms in a drinking water supply line by enteric bacteria
Author(s) -
Robinson P.J.,
Walker J.T.,
Keevil C.W.,
Cole J.
Publication year - 1995
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1995.tb07577.x
Subject(s) - biofilm , chemostat , reporter gene , bacteria , microbiology and biotechnology , escherichia coli , green fluorescent protein , plasmid , biology , bioreporter , lac operon , fluorescence , fluorescence microscope , gene , chemistry , gene expression , biochemistry , genetics , physics , quantum mechanics
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 β‐galactosidase from pnirB .