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 -