Internal arsenite bioassay calibration using multiple reporter cell lines

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State: Public
Version: Final published version
Serval ID
serval:BIB_D3D7C035CF00
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Internal arsenite bioassay calibration using multiple reporter cell lines
Journal
Microbial Biotechnology
Author(s)
Wackwitz A., Harms H., Chatzinotas A., Breuer U., Vogne C., van der Meer J. R.
ISSN
1751-7915
Publication state
Published
Issued date
2008
Peer-reviewed
Oui
Volume
1
Number
2
Pages
149-157
Language
english
Abstract
Bioassays with bioreporter bacteria are usually calibrated with analyte solutions of known concentrations that are analysed along with the samples of interest. This is done as bioreporter output (the intensity of light, fluorescence or colour) does not only depend on the target concentration, but also on the incubation time and physiological activity of the cells in the assay. Comparing the bioreporter output with standardized colour tables in the field seems rather difficult and error-prone. A new approach to control assay variations and improve application ease could be an internal calibration based on the use of multiple bioreporter cell lines with drastically different reporter protein outputs at a given analyte concentration. To test this concept, different Escherichia coli-based bioreporter strains expressing either cytochrome c peroxidase (CCP, or CCP mutants) or β-galactosidase upon induction with arsenite were constructed. The reporter strains differed either in the catalytic activity of the reporter protein (for CCP) or in the rates of reporter protein synthesis (for β-galactosidase), which, indeed, resulted in output signals with different intensities at the same arsenite concentration. Hence, it was possible to use combinations of these cell lines to define arsenite concentration ranges at which none, one or more cell lines gave qualitative (yes/no) visible signals that were relatively independent of incubation time or bioreporter activity. The discriminated concentration ranges would fit very well with the current permissive (e.g. World Health Organization) levels of arsenite in drinking water (10 µg l−1).
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Create date
17/02/2009 16:39
Last modification date
20/08/2019 15:53
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