Details
| Original language | English |
|---|---|
| Pages (from-to) | 2-8 |
| Number of pages | 7 |
| Journal | Sensors and Actuators, B: Chemical |
| Volume | 90 |
| Issue number | 1-3 |
| Publication status | Published - 26 Feb 2003 |
| Event | 6th european Conference on Optical Chemical - Manchester, United Kingdom (UK) Duration: 7 Apr 2002 → 7 Apr 2002 |
Abstract
Genetically modified bacteria, engineered to generate a quantifiable signal in response to pre-determined sets of environmental conditions, may serve as combined sensing/reporting elements in whole-cell biosensors. We have compared two of the several available reporter genes in such cells: green fluorescent proteins (GFPs) (Aquorea victoria gfp) and bioluminescence (Vibrio fischeri luxCDABE) genes, fused to either SOS (recA) or heat shock (grpE) promoters. In both cases, bacterial bioluminescence allowed faster and more sensitive detection of the model toxicants; the fluorescent reporter proteins were much more stable, and following long-term exposure allowed detection at levels similar to that of the bioluminescent sensors. From the two green fluorescent proteins tested, enhanced GFP (EGFP) displayed a more rapid response and higher signal intensity than GFPuv. To combine the advantages of both reporter functions, representatives of both types were jointly encapsulated in a sol-gel matrix and immobilized onto a glass surface, to generate a bioluminescent toxicity and a fluorescent genotoxicity sensor. The dual-function sensor detected both toxic and genotoxic model compounds with no interference from the co-immobilized member.
Keywords
- Bioluminescence, Escherichia coli, Fluorescence, Genotoxicity, Green fluorescent protein, Sol-gel, Toxicity
ASJC Scopus subject areas
- Materials Science(all)
- Electronic, Optical and Magnetic Materials
- Physics and Astronomy(all)
- Instrumentation
- Physics and Astronomy(all)
- Condensed Matter Physics
- Materials Science(all)
- Surfaces, Coatings and Films
- Materials Science(all)
- Metals and Alloys
- Engineering(all)
- Electrical and Electronic Engineering
- Materials Science(all)
- Materials Chemistry
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In: Sensors and Actuators, B: Chemical, Vol. 90, No. 1-3, 26.02.2003, p. 2-8.
Research output: Contribution to journal › Conference article › Research › peer review
}
TY - JOUR
T1 - Fluorescence and bioluminescence reporter functions in genetically modified bacterial sensor strains
AU - Sagi, Eran
AU - Hever, Navit
AU - Rosen, Rachel
AU - Bartolone, Amelita J.
AU - Premkumar, J. Rajan
AU - Ulber, Roland
AU - Lev, Ovadia
AU - Scheper, Thomas
AU - Belkin, Shimshon
N1 - Funding information: Research was supported by DARPA grant number N00173-01-1-G009, by a research grant from the Land Niedersachen Min. f. Wissenchaft & Kultur (Germany) 16 November 1998—25 A.5—76 251-99-2/98 (ZN549) and by the Israeli Ministry of Science, Culture and Sport (Infrastructure grant 1319-1-98 and the France–Israel Cooperative Research Program). We are grateful to U. Alon, T. Van Dyk and R.A. LaRossa for gifts of strains and plasmids.
PY - 2003/2/26
Y1 - 2003/2/26
N2 - Genetically modified bacteria, engineered to generate a quantifiable signal in response to pre-determined sets of environmental conditions, may serve as combined sensing/reporting elements in whole-cell biosensors. We have compared two of the several available reporter genes in such cells: green fluorescent proteins (GFPs) (Aquorea victoria gfp) and bioluminescence (Vibrio fischeri luxCDABE) genes, fused to either SOS (recA) or heat shock (grpE) promoters. In both cases, bacterial bioluminescence allowed faster and more sensitive detection of the model toxicants; the fluorescent reporter proteins were much more stable, and following long-term exposure allowed detection at levels similar to that of the bioluminescent sensors. From the two green fluorescent proteins tested, enhanced GFP (EGFP) displayed a more rapid response and higher signal intensity than GFPuv. To combine the advantages of both reporter functions, representatives of both types were jointly encapsulated in a sol-gel matrix and immobilized onto a glass surface, to generate a bioluminescent toxicity and a fluorescent genotoxicity sensor. The dual-function sensor detected both toxic and genotoxic model compounds with no interference from the co-immobilized member.
AB - Genetically modified bacteria, engineered to generate a quantifiable signal in response to pre-determined sets of environmental conditions, may serve as combined sensing/reporting elements in whole-cell biosensors. We have compared two of the several available reporter genes in such cells: green fluorescent proteins (GFPs) (Aquorea victoria gfp) and bioluminescence (Vibrio fischeri luxCDABE) genes, fused to either SOS (recA) or heat shock (grpE) promoters. In both cases, bacterial bioluminescence allowed faster and more sensitive detection of the model toxicants; the fluorescent reporter proteins were much more stable, and following long-term exposure allowed detection at levels similar to that of the bioluminescent sensors. From the two green fluorescent proteins tested, enhanced GFP (EGFP) displayed a more rapid response and higher signal intensity than GFPuv. To combine the advantages of both reporter functions, representatives of both types were jointly encapsulated in a sol-gel matrix and immobilized onto a glass surface, to generate a bioluminescent toxicity and a fluorescent genotoxicity sensor. The dual-function sensor detected both toxic and genotoxic model compounds with no interference from the co-immobilized member.
KW - Bioluminescence
KW - Escherichia coli
KW - Fluorescence
KW - Genotoxicity
KW - Green fluorescent protein
KW - Sol-gel
KW - Toxicity
UR - http://www.scopus.com/inward/record.url?scp=0037457509&partnerID=8YFLogxK
U2 - 10.1016/S0925-4005(03)00014-5
DO - 10.1016/S0925-4005(03)00014-5
M3 - Conference article
AN - SCOPUS:0037457509
VL - 90
SP - 2
EP - 8
JO - Sensors and Actuators, B: Chemical
JF - Sensors and Actuators, B: Chemical
SN - 0925-4005
IS - 1-3
T2 - 6th european Conference on Optical Chemical
Y2 - 7 April 2002 through 7 April 2002
ER -