Open AccessConstruction of a modified mini‐Tn 5 lux CDABE transposon for the development of bacterial biosensors for ecotoxicity testing
Author(s) -
Weitz Hedda J,
Ritchie Jenny M,
Bailey David A,
Horsburgh Alison M,
Killham Ken,
Glover L.Anne
Publication year - 2001
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.1574-6968.2001.tb10598.x
Subject(s) - bioreporter , biology , pseudomonas putida , bioluminescence , ecotoxicity , pseudomonas fluorescens , pseudomonas , transposable element , transposon mutagenesis , microbiology and biotechnology , reporter gene , bacteria , toxicity , genetics , mutant , ecology , gene expression , chemistry , organic chemistry , gene
A mini‐Tn 5 transposon was modified to introduce a promoterless lux CDABE cassette from Vibrio fischeri into environmentally relevant bacterial strains in order to develop bioluminescence‐based biosensors for toxicity testing. The mini‐Tn 5 lux CDABE 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 Tn 5 lux CDABE and P. putida F1 Tn 5 lux CDABE 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.