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Response Characteristics of Arsenic-Sensitive Bioreporters Expressing the gfp Reporter Gene
Authors:Mona Wells  Michael Gösch  Hauke Harms  Jan Roelof van der Meer
Affiliation:Swiss Federal Institute of Technology Lausanne (EPFL), Institute of Environmental Science and Technology, CH-1015, Lausanne, Switzerland
Department of Applied Optics, Swiss Federal Institute of Technology Lausanne (EPFL), CH-1015, Lausanne, Switzerland
Department of Environmental Microbiology, UFZ Centre for Environmental Research Leipzig-Halle, Permoserstra?e 15, D-04318, Leipzig, Germany
Department of Fundamental Microbiology, University of Lausanne, CH-1015, Lausanne, Switzerland
Abstract:
This paper describes the development of an analytical technique for arsenic analyses that is based on genetically-modified bioreporter bacteria bearing a gene encoding for the production of a green fluorescent protein (gfp). Upon exposure to arsenic (in the aqueous form of arsenite), the bioreporter production of the fluorescent reporter molecule is monitored spectroscopically. We compared the response measured as a function of time and concentration by steady-state fluorimetry (SSF) to that measured by epi-fluorescent microscopy (EFM). SSF is a bulk technique; as such it inherently yields less information, whereas EFM monitors the response of many individual cells simultaneously and data can be processed in terms of population averages or subpopulations. For the bioreporter strain used here, as well as for the literature we cite, the two techniques exhibit similar performance characteristics. The results presented here show that the EFM technique can compete with SSF and shows substantially more promise for future improvement; it is a matter of research interest to develop optimized methods of EFM image analysis and statistical data treatment. EFM is a conduit for understanding the dynamics of individual cell response vs. population response, which is not only a matter of research interest, but is also promising in the practical terms of developing micro-scale analysis.
Keywords:: E. coli DH5α     gfp   bioreporter   arsenic   epi-fluorescent microscopy   fluorimetry.
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