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

• Published in: FEMS microbiology letters Vol. 197; no. 2; pp. 159 - 165
• Main Authors: Weitz, Hedda J., Ritchie, Jenny M., Bailey, David A., Horsburgh, Alison M., Killham, Ken, Glover, L. Anne
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Publishing Ltd 13.04.2001; Blackwell; Oxford University Press
• Subjects: Bacteria; Bacteria - genetics; Bacteria - growth & development; Bacteriological methods and techniques used in bacteriology; Bacteriology; Bioassays; Biological and medical sciences; Bioluminescence; Biosensing Techniques; Biosensor; Biosensors; Chlorophenols - analysis; Cloning, Molecular; Conjugation, Genetic; Copper - analysis; DNA Transposable Elements - genetics; Fundamental and applied biological sciences. Psychology; Genetics; Groundwater; Growth rate; Luminescent Measurements; luxCDABE; luxCDABE gene; Microbiology; Mini‐Tn5; Pseudomonas - genetics; Pseudomonas - growth & development; Pseudomonas fluorescens; Pseudomonas putida; Strains (organisms); Toxicity; Toxicity testing; Toxicity Tests; transposon mini-TN5; Transposons; Vibrio - genetics; Vibrio fischeri; Water - chemistry; Water analysis; Water sampling; Waterborne diseases; Zinc - analysis; Bioluminescence; CDABE; Mini-Tn; Toxicity; Biosensor; Pseudomonadales; Ecotoxicology; Growth rate; Growth; Transposon; Pseudomonas fluorescens; Standards; Strain; Promoter; Vibrio fischeri; Development; Pseudomonadaceae; Vibrionaceae; Bacteria; Pseudomonas putida
• ISSN: 0378-1097; 1574-6968
• DOI: 10.1111/j.1574-6968.2001.tb10598.x

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.