Fluorescence and bioluminescence reporter functions in genetically modified bacterial sensor strains

Eran Sagi, Navit Hever, Rachel Rosen, Amelita J. Bartolone, J. Rajan Premkumar, Roland Ulber, Ovadia Lev, Thomas Scheper, Shimshon Belkin*

*Corresponding author for this work

Research output: Contribution to journalConference articlepeer-review

87 Scopus citations

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.

Original languageAmerican English
Pages (from-to)2-8
Number of pages7
JournalSensors and Actuators, B: Chemical
Volume90
Issue number1-3
DOIs
StatePublished - 20 Apr 2003
EventProceedingsof the 6th european Conference on Optical Chemical - Manchester, United Kingdom
Duration: 7 Apr 20027 Apr 2002

Bibliographical note

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.

Keywords

  • Bioluminescence
  • Escherichia coli
  • Fluorescence
  • Genotoxicity
  • Green fluorescent protein
  • Sol-gel
  • Toxicity

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