Journal
ACS SYNTHETIC BIOLOGY
Volume 8, Issue 9, Pages 1968-1975Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acssynbio.9b00144
Keywords
Saccharomyces cerevisiae; promoter engineering; biosensor; maximum dynamic range; malonyl-CoA; fapO/FapR
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Funding
- Novo Nordisk Foundation [NNF1OCC1016517]
- Angpanneforeningen's Foundation for Research and Development
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Metabolite biosensors are useful tools for high throughput screening approaches and pathway regulation approaches. An important feature of biosensors is the dynamic range. To expand the maximum dynamic range of a transcription factor-based biosensor in Saccharomyces cerevi-siae, using the fapO/FapR system from Bacillus subtilis as an example case, five native promoters, including constitutive and glucose-regulated ones, were modified. By evaluating different binding site (BS) positions in the core promoters, we identified locations that resulted in a high maximum dynamic range with low expression under repressed conditions. We further identified BS positions in the upstream element region of the TEF1 promoter that did not influence the native promoter strength but resulted in repression in the presence of a chimeric repressor consisting of FapR and the yeast repressor Mig1. These modified promoters with broad dynamic ranges will provide useful information for the engineering of future biosensors and their use in complex genetic circuits.
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