期刊
ACS SYNTHETIC BIOLOGY
卷 9, 期 1, 页码 84-94出版社
AMER CHEMICAL SOC
DOI: 10.1021/acssynbio.9b00348
关键词
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资金
- University of Wisconsin Madison
- Army Research Office [W911NF1710043]
- Air Force Research Laboratory Center of Excellence Grant [FA8650-15-2-5518]
- DARPA 1000 Molecules Program [HR0011-1S-C-0084]
- David and Lucile Packard Foundation
- Camille Dreyfus Teacher-Scholar Program
- Searle Funds at the Chicago Community Trust
- National Institutes of Health Training Grant through Northwestern University's Biotechnology Training Program [T32GM008449]
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.
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