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 journalArticle

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 languageEnglish
Pages (from-to)159-165
Number of pages6
JournalFEMS Microbiology Letters
Volume197
DOIs
Publication statusPublished - 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

Cite this

Construction of a modified mini-Tn5 luxCDABE transposen for the development of bacterial biosensors for ecotoxicity testing. / Weitz, Hedda; Ritchie, J. M.; Bailey, D. A.; Horsburgh, Alison Margaret; Killham, Kenneth Stuart; Glover, Lesley Anne.

In: FEMS Microbiology Letters, Vol. 197, 2001, p. 159-165.

Research output: Contribution to journalArticle

Weitz, Hedda ; Ritchie, J. M. ; Bailey, D. A. ; Horsburgh, Alison Margaret ; Killham, Kenneth Stuart ; Glover, Lesley Anne. / Construction of a modified mini-Tn5 luxCDABE transposen for the development of bacterial biosensors for ecotoxicity testing. In: FEMS Microbiology Letters. 2001 ; Vol. 197. pp. 159-165.
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