Construction of a whole-cell gene reporter for the fluorescent bioassay of nitrate

Taylor, Clare J., Bain, Lindsey A., Richardson, David J. ORCID:, Spiro, Stephen and Russell, David A. (2004) Construction of a whole-cell gene reporter for the fluorescent bioassay of nitrate. Analytical Biochemistry, 328 (1). pp. 60-66. ISSN 1096-0309

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The development of a whole-cell fluorescence-based biosensor for nitrate is reported. The sensor is Escherichia coli transformed with a plasmid (pPNARGFP) in which the promoter and regulatory regions of the membrane-bound nitrate reductase narGHJI operon (Pnar) are fused to a gfp gene encoding green fluorescent protein (GFP). Pnar-gfp activity was measured at a range of nitrate concentrations using whole-cell GFP fluorescence. The bioassay conditions have been optimized so that the fluorescence intensity is proportional to the extracellular nitrate concentration. The developed bioassay has established that E. coli (pPNARGFP) can be used for the quantitative determination of nitrate in environmental waters without interference from other electron acceptors, e.g., nitrite, dimethyl sulfoxide, trimethylamine-N-oxide and fumerate, and azide, an inhibitor of redox-active proteins.

Item Type: Article
Faculty \ School: Faculty of Science > School of Biological Sciences
Faculty of Science > School of Chemistry
Depositing User: Rachel Smith
Date Deposited: 24 Jan 2011 11:59
Last Modified: 16 May 2023 08:30
DOI: 10.1016/j.ab.2004.01.013

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