Tunable reporter signal production in feedback-uncoupled arsenic bioreporters.

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Serval ID
serval:BIB_5B9F1130187F
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Tunable reporter signal production in feedback-uncoupled arsenic bioreporters.
Journal
Microbial Biotechnology
Author(s)
Merulla D., Hatzimanikatis V., van der Meer J.R.
ISSN
1751-7915 (Electronic)
ISSN-L
1751-7915
Publication state
Published
Issued date
2013
Peer-reviewed
Oui
Volume
6
Number
5
Pages
503-514
Language
english
Notes
Publication types: Journal Article Publication Status: ppublish
Abstract
Escherichia coli-based bioreporters for arsenic detection are typically based on the natural feedback loop that controls ars operon transcription. Feedback loops are known to show a wide range linear response to the detriment of the overall amplification of the incoming signal. While being a favourable feature in controlling arsenic detoxification for the cell, a feedback loop is not necessarily the most optimal for obtaining highest sensitivity and response in a designed cellular reporter for arsenic detection. Here we systematically explore the effects of uncoupling the topology of arsenic sensing circuitry on the developed reporter signal as a function of arsenite concentration input. A model was developed to describe relative ArsR and GFP levels in feedback and uncoupled circuitry, which was used to explore new ArsR-based synthetic circuits. The expression of arsR was then placed under the control of a series of constitutive promoters, which differed in promoter strength, and which could be further modulated by TetR repression. Expression of the reporter gene was maintained under the ArsR-controlled Pars promoter. ArsR expression in the systems was measured by using ArsR-mCherry fusion proteins. We find that stronger constitutive ArsR production decreases arsenite-dependent EGFP output from Pars and vice versa. This leads to a tunable series of arsenite-dependent EGFP outputs in a variety of systematically characterized circuitries. The higher expression levels and sensitivities of the response curves in the uncoupled circuits may be useful for improving field-test assays using arsenic bioreporters.
Pubmed
Web of science
Open Access
Yes
Create date
08/09/2013 9:12
Last modification date
20/08/2019 14:14
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