Reporter genes lucFF, luxCDABE, gfp, and dsred have different characteristics in whole-cell bacterial sensors.
The selection of a genetic reporter can be difficult because of the wide range of genes available. In order to reduce the selection, we compared the performance of different reporter genes: firefly luciferase (Photinus pyralis lucFF), bacterial luciferase operon (Photorhabdus luminescens luxCDABE), green fluorescent protein (Aequorea victoria gfp), and red fluorescent protein (Discosoma sp. dsred) in whole-cell bacterial sensors. Escherichia coli sensor bacteria were engineered to contain a reporter plasmid that carries the reporter gene under the control of mercury- (mer from Tn21) or arsenite- (ars from R773) responsive regulatory units. Characteristics of the strains were studied by using different arsenite or mercury concentrations and incubation times. The lowest detectable concentration of analytes and the fastest responses were achieved with lucFF or luxCDABE as reporter genes. The fluorescent proteins, GFP and DsRed, gave responses at higher analyte concentrations and after significantly longer incubation times. The results indicate that luciferases are better reporters in whole-cell sensor bacteria.
Hakkila
Kaisa
K
Department of Biotechnology, University of Turku, Turku, Finland. kaihak@utu.fi
Maksimow
Mikael
M
Karp
Matti
M
Virta
Marko
M
D000818:Animals; D018053:Arsenites / Q000494:pharmacology; D015374:Biosensing Techniques / Q000379:methods*; D001517:Coleoptera / Q000201:enzymology; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005809:Genes, Regulator / Q000187:drug effects; D017930:Genes, Reporter / Q000235:genetics; D049452:Green Fluorescent Proteins; D008156:Luciferases / Q000235:genetics*; D008164:Luminescent Proteins / Q000187:drug effects / Q000235:genetics; D008628:Mercury / Q000494:pharmacology; D000097573:Red Fluorescent Protein
D018053:Arsenites; D049452:Green Fluorescent Proteins; D008156:Luciferases; D008164:Luminescent Proteins; D008628:Mercury; D000097573:Red Fluorescent Protein; C071832:alkanol monooxygenase (FMN-linked); C015001:arsenite