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Reporter genes and fluorescent probes for studying the colonisation of biofilms in a drinking water supply line by enteric bacteria

Reporter genes and fluorescent probes for studying the colonisation of biofilms in a drinking water supply line by enteric bacteria
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 β-galactosidase from pnirB.

Anaerobic regulation, Biofilm, Differential interference contrast microscopy, Escherichia coli, Escherichia coli nirB promoter, Fluorescence microscopy, gusA reporter gene, lacZ reporter gene
0378-1097
183-188
Robinson, P. J.
447c51f6-f710-40c7-bce9-2caf7c4876f8
Walker, J. T.
2bb5ed4e-d929-47e4-97ba-70641716acd7
Keevil, C. W.
cb7de0a7-ce33-4cfa-af52-07f99e5650eb
Cole, J.
7ba340f7-acf6-4a25-9e82-81823654dbdf
Robinson, P. J.
447c51f6-f710-40c7-bce9-2caf7c4876f8
Walker, J. T.
2bb5ed4e-d929-47e4-97ba-70641716acd7
Keevil, C. W.
cb7de0a7-ce33-4cfa-af52-07f99e5650eb
Cole, J.
7ba340f7-acf6-4a25-9e82-81823654dbdf

Robinson, P. J., Walker, J. T., Keevil, C. W. and Cole, J. (1995) Reporter genes and fluorescent probes for studying the colonisation of biofilms in a drinking water supply line by enteric bacteria. FEMS Microbiology Letters, 129 (2-3), 183-188. (doi:10.1016/0378-1097(95)00155-X).

Record type: Article

Abstract

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.

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More information

Accepted/In Press date: 14 April 1995
e-pub ahead of print date: 1 June 1995
Published date: 15 June 1995
Keywords: Anaerobic regulation, Biofilm, Differential interference contrast microscopy, Escherichia coli, Escherichia coli nirB promoter, Fluorescence microscopy, gusA reporter gene, lacZ reporter gene

Identifiers

Local EPrints ID: 431342
URI: http://eprints.soton.ac.uk/id/eprint/431342
ISSN: 0378-1097
PURE UUID: d3ddc369-b55d-4714-90c1-f03c4462d8e3
ORCID for C. W. Keevil: ORCID iD orcid.org/0000-0003-1917-7706

Catalogue record

Date deposited: 29 May 2019 16:30
Last modified: 30 May 2019 00:36

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Contributors

Author: P. J. Robinson
Author: J. T. Walker
Author: C. W. Keevil ORCID iD
Author: J. Cole

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