Tunable reporter signal production in feedback-uncoupled arsenic bioreporters.

Détails

Ressource 1Télécharger: BIB_5B9F1130187F.P001.pdf (1336.66 [Ko])
Etat: Public
Version: de l'auteur⸱e
ID Serval
serval:BIB_5B9F1130187F
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
Tunable reporter signal production in feedback-uncoupled arsenic bioreporters.
Périodique
Microbial Biotechnology
Auteur⸱e⸱s
Merulla D., Hatzimanikatis V., van der Meer J.R.
ISSN
1751-7915 (Electronic)
ISSN-L
1751-7915
Statut éditorial
Publié
Date de publication
2013
Peer-reviewed
Oui
Volume
6
Numéro
5
Pages
503-514
Langue
anglais
Notes
Publication types: Journal Article Publication Status: ppublish
Résumé
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
Oui
Création de la notice
08/09/2013 9:12
Dernière modification de la notice
20/08/2019 14:14
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