Construction of a modified mini-Tn5 luxCDABE transposen for the development of bacterial biosensors for ecotoxicity testing

Hedda Weitz; J. M. Ritchie; D. A. Bailey; Alison Margaret Horsburgh; Kenneth Stuart Killham; Lesley Anne Glover

doi:10.1111/j.1574-6968.2001.tb10598.x

Construction of a modified mini-Tn5 luxCDABE transposen for the development of bacterial biosensors for ecotoxicity testing

Hedda Weitz, J. M. Ritchie, D. A. Bailey, Alison Margaret Horsburgh, Kenneth Stuart Killham, Lesley Anne Glover

Research output: Contribution to journal › Article › peer-review

Abstract

A mini-Tn5 transposon was modified to introduce a promoterless luxCDABE cassette from Vibrio fischeri into environmentally relevant bacterial strains in order to develop bioluminescence-based biosensors for toxicity testing. The mini-Tn5 luxCDABE transposon was chromosomally integrated downstream from an active promoter into two Pseudomonas strains (Pseudomonas fluorescens 8866 and Pseudomonas putida F1). Characterisation of the bioluminescent transconjugants demonstrated that the transposon integration was stable and had no effect on growth rate. Both P. fluorescens 8866 Tn5 luxCDABE and P. putida F1 Tn5 luxCDABE were used to assess the toxicity of standard solutions (Cu, Zn and 3,5-DCP) as well as Cu- and 3,5-DCP-spiked groundwater samples. They were successfully used for bioluminescence-based bioassays and the potential value of using different bacterial biosensors for ecotoxicity testing was shown. (C) 2001 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.

Original language	English
Pages (from-to)	159-165
Number of pages	6
Journal	FEMS Microbiology Letters
Volume	197
DOIs	https://doi.org/10.1111/j.1574-6968.2001.tb10598.x
Publication status	Published - 2001

Keywords

luxCDABE
mini-Tn5
bioluminescence
biosensor
toxicity
GRAM-NEGATIVE BACTERIA
PSEUDOMONAS-FLUORESCENS
BIOLUMINESCENT REPORTER
CHROMOSOMAL INSERTION
TOXICITY ASSESSMENT
HEAVY-METALS
LUX
GENES
MUTAGENESIS
TOLUENE

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10.1111/j.1574-6968.2001.tb10598.x

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title = "Construction of a modified mini-Tn5 luxCDABE transposen for the development of bacterial biosensors for ecotoxicity testing",

abstract = "A mini-Tn5 transposon was modified to introduce a promoterless luxCDABE cassette from Vibrio fischeri into environmentally relevant bacterial strains in order to develop bioluminescence-based biosensors for toxicity testing. The mini-Tn5 luxCDABE transposon was chromosomally integrated downstream from an active promoter into two Pseudomonas strains (Pseudomonas fluorescens 8866 and Pseudomonas putida F1). Characterisation of the bioluminescent transconjugants demonstrated that the transposon integration was stable and had no effect on growth rate. Both P. fluorescens 8866 Tn5 luxCDABE and P. putida F1 Tn5 luxCDABE were used to assess the toxicity of standard solutions (Cu, Zn and 3,5-DCP) as well as Cu- and 3,5-DCP-spiked groundwater samples. They were successfully used for bioluminescence-based bioassays and the potential value of using different bacterial biosensors for ecotoxicity testing was shown. (C) 2001 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.",

keywords = "luxCDABE, mini-Tn5, bioluminescence, biosensor, toxicity, GRAM-NEGATIVE BACTERIA, PSEUDOMONAS-FLUORESCENS, BIOLUMINESCENT REPORTER, CHROMOSOMAL INSERTION, TOXICITY ASSESSMENT, HEAVY-METALS, LUX, GENES, MUTAGENESIS, TOLUENE",

author = "Hedda Weitz and Ritchie, {J. M.} and Bailey, {D. A.} and Horsburgh, {Alison Margaret} and Killham, {Kenneth Stuart} and Glover, {Lesley Anne}",

year = "2001",

doi = "10.1111/j.1574-6968.2001.tb10598.x",

language = "English",

volume = "197",

pages = "159--165",

journal = "FEMS Microbiology Letters",

issn = "0378-1097",

publisher = "Oxford University Press",

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T1 - Construction of a modified mini-Tn5 luxCDABE transposen for the development of bacterial biosensors for ecotoxicity testing

AU - Weitz, Hedda

AU - Ritchie, J. M.

AU - Bailey, D. A.

AU - Horsburgh, Alison Margaret

AU - Killham, Kenneth Stuart

AU - Glover, Lesley Anne

PY - 2001

Y1 - 2001

N2 - A mini-Tn5 transposon was modified to introduce a promoterless luxCDABE cassette from Vibrio fischeri into environmentally relevant bacterial strains in order to develop bioluminescence-based biosensors for toxicity testing. The mini-Tn5 luxCDABE transposon was chromosomally integrated downstream from an active promoter into two Pseudomonas strains (Pseudomonas fluorescens 8866 and Pseudomonas putida F1). Characterisation of the bioluminescent transconjugants demonstrated that the transposon integration was stable and had no effect on growth rate. Both P. fluorescens 8866 Tn5 luxCDABE and P. putida F1 Tn5 luxCDABE were used to assess the toxicity of standard solutions (Cu, Zn and 3,5-DCP) as well as Cu- and 3,5-DCP-spiked groundwater samples. They were successfully used for bioluminescence-based bioassays and the potential value of using different bacterial biosensors for ecotoxicity testing was shown. (C) 2001 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.

AB - A mini-Tn5 transposon was modified to introduce a promoterless luxCDABE cassette from Vibrio fischeri into environmentally relevant bacterial strains in order to develop bioluminescence-based biosensors for toxicity testing. The mini-Tn5 luxCDABE transposon was chromosomally integrated downstream from an active promoter into two Pseudomonas strains (Pseudomonas fluorescens 8866 and Pseudomonas putida F1). Characterisation of the bioluminescent transconjugants demonstrated that the transposon integration was stable and had no effect on growth rate. Both P. fluorescens 8866 Tn5 luxCDABE and P. putida F1 Tn5 luxCDABE were used to assess the toxicity of standard solutions (Cu, Zn and 3,5-DCP) as well as Cu- and 3,5-DCP-spiked groundwater samples. They were successfully used for bioluminescence-based bioassays and the potential value of using different bacterial biosensors for ecotoxicity testing was shown. (C) 2001 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.

KW - luxCDABE

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KW - biosensor

KW - toxicity

KW - GRAM-NEGATIVE BACTERIA

KW - PSEUDOMONAS-FLUORESCENS

KW - BIOLUMINESCENT REPORTER

KW - CHROMOSOMAL INSERTION

KW - TOXICITY ASSESSMENT

KW - HEAVY-METALS

KW - LUX

KW - GENES

KW - MUTAGENESIS

KW - TOLUENE

U2 - 10.1111/j.1574-6968.2001.tb10598.x

DO - 10.1111/j.1574-6968.2001.tb10598.x

M3 - Article

SN - 0378-1097

VL - 197

SP - 159

EP - 165

JO - FEMS Microbiology Letters

JF - FEMS Microbiology Letters

ER -

Construction of a modified mini-Tn5 luxCDABE transposen for the development of bacterial biosensors for ecotoxicity testing

Abstract

Keywords

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