title: Comparison of three genetically modified Escherichia coli biosensor strains for amperometric tetracycline measurement.
abstract: Three separate genetic strategies, based upon the induced expression of three different genes (lacZ, selA and nuoA) were tested to provide the SciTox assay with sensitive and specific detection of the antibiotic tetracycline (Tet). All three strategies relied on gene induction from the Tn10 tetA promoter. Both lacZ and nuoA biosensors responded specifically and sensitively to sub-inhibitory concentrations of Tet. However, the selA-based assay was not sensitive enough to detect Tet in the SciTox assay. The detection limits for Tet of the lacZ and nuoA biosensor strains were 0.11 μg ml(-1) and 0.0026 μg ml(-1), respectively, and their linear ranges were 0.1-1 μg ml(-1) and 0-0.01 μg ml(-1), respectively. While lacZ has previously been used as a reporter gene in an amperometric bioassay, nuoA is a novel and more sensitive reporter gene. This is the first report in which a respiratory gene was used as a reporter gene in an amperometric biosensor. The results indicate that this approach can produce a highly sensitive detection system. In order to test whether the new system could be used to detect other chemicals, the nuoA gene was re-engineered to be driven by the copper-inducible copA promoter. Using this strain, the SciTox assay was found to be able to specifically detect copper and silver ions.
authors.affiliation: Lincoln University, P.O. Box 84, Lincoln University, Lincoln 7647, New Zealand. Electronic address: Songw2@hotmail.com.
authors.forename: Wenfeng
authors.initials: W
authors.lastname: Song
authors.affiliation: Lincoln Ventures Limited, P.O. Box 133, Lincoln University, Lincoln 7647, New Zealand. Electronic address: pascon@lvl.co.nz.
authors.forename: Neil
authors.initials: N
authors.lastname: Pasco
authors.affiliation: Lincoln University, P.O. Box 84, Lincoln University, Lincoln 7647, New Zealand. Electronic address: Ravi.Gooneratne@lincoln.ac.nz.
authors.forename: Ravi
authors.initials: R
authors.lastname: Gooneratne
authors.affiliation: Lincoln Ventures Limited, P.O. Box 133, Lincoln University, Lincoln 7647, New Zealand. Electronic address: weldr@lvl.co.nz.
authors.forename: Richard J
authors.initials: RJ
authors.lastname: Weld
mesh_terms: D017920:Antiporters / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics; D001483:Base Sequence; D015374:Biosensing Techniques / Q000379:methods*; D004269:DNA, Bacterial / Q000235:genetics; D055664:Electrochemical Techniques; D042967:Electron Transport Complex I / Q000235:genetics; D004926:Escherichia coli / Q000187:drug effects* / Q000235:genetics* / Q000378:metabolism; D005798:Genes, Bacterial / Q000187:drug effects; D017930:Genes, Reporter / Q000187:drug effects; D005818:Genetic Engineering; D007763:Lac Operon / Q000187:drug effects; D011401:Promoter Regions, Genetic; D013752:Tetracycline / Q000032:analysis* / Q000633:toxicity; D014166:Transferases / Q000235:genetics; D014874:Water Pollutants, Chemical / Q000032:analysis / Q000633:toxicity
chemical_list: D017920:Antiporters; D001426:Bacterial Proteins; D004269:DNA, Bacterial; D014874:Water Pollutants, Chemical; C079051:tetA protein, Bacteria; D014166:Transferases; C060600:selenium transferase; D042967:Electron Transport Complex I; D013752:Tetracycline