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Design of a Transcriptional Biosensor for the Portable, On-Demand Detection of Cyanuric Acid.
- Source :
-
ACS synthetic biology [ACS Synth Biol] 2020 Jan 17; Vol. 9 (1), pp. 84-94. Date of Electronic Publication: 2019 Dec 26. - Publication Year :
- 2020
-
Abstract
- Rapid molecular biosensing is an emerging application area for synthetic biology. Here, we engineer a portable biosensor for cyanuric acid (CYA), an analyte of interest for human and environmental health, using a LysR-type transcription regulator (LTTR) from Pseudomonas within the context of Escherichia coli gene expression machinery. To overcome cross-host portability challenges of LTTRs, we rationally engineered hybrid Pseudomonas-E. coli promoters by integrating DNA elements required for transcriptional activity and ligand-dependent regulation from both hosts, which enabled E. coli to function as a whole-cell biosensor for CYA. To alleviate challenges of whole-cell biosensing, we adapted these promoter designs to function within a freeze-dried E. coli cell-free system to sense CYA. This portable, on-demand system robustly detects CYA within an hour from laboratory and real-world samples and works with both fluorescent and colorimetric reporters. This work elucidates general principles to facilitate the engineering of a wider array of LTTR-based environmental sensors.
- Subjects :
- Bacterial Proteins metabolism
Cell-Free System
Chimera genetics
Gene Expression
Gene Expression Regulation, Bacterial
Ligands
Plasmids genetics
Promoter Regions, Genetic
Synthetic Biology methods
Transcription Factors metabolism
Biosensing Techniques methods
Escherichia coli genetics
Pseudomonas genetics
Transcription, Genetic
Triazines analysis
Subjects
Details
- Language :
- English
- ISSN :
- 2161-5063
- Volume :
- 9
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- ACS synthetic biology
- Publication Type :
- Academic Journal
- Accession number :
- 31825601
- Full Text :
- https://doi.org/10.1021/acssynbio.9b00348