Abstract
Here we describe the engineering of a distant homolog of the Tet repressor, CamR, isolated from Pseudomonas putida, that is regulated by camphor, a very inexpensive small molecule (at micromolar concentrations) for use in Saccharomyces cerevisiae. The repressor was engineered by expression from a constitutive yeast promoter, fusion to a viral activator protein cassette, and codon optimization. A suitable promoter responsive to the CamR fusion protein was engineered by embedding a P. putida operator binding sequence within an upstream activating sequence (UAS)-less CYC1 promoter from S. cerevisiae. The switch, named the Camphor-Off switch, activates expression of a reporter gene in camphor-free media and represses it with micromolar concentrations of camphor.
Original language | English (US) |
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Pages (from-to) | 1983-1990 |
Number of pages | 8 |
Journal | G3: Genes, Genomes, Genetics |
Volume | 5 |
Issue number | 10 |
DOIs | |
State | Published - 2015 |
Keywords
- CamR
- Pseudomonas putida
- Tet system
- TetR homolog
ASJC Scopus subject areas
- Molecular Biology
- Genetics
- Genetics(clinical)