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Development of a Tightly Controlled Off Switch for Saccharomyces cerevisiae Regulated by Camphor, a Low-Cost Natural Product.

Ikushima S, Zhao Y, Boeke JD - G3 (Bethesda) (2015)

Bottom Line: 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.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.

View Article: PubMed Central - PubMed

Affiliation: High Throughput Biology Center and Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205 Central Laboratories for Key Technologies, KIRIN Company Limited, Yokohama, Kanagawa 236-0004, Japan.

No MeSH data available.


Related in: MedlinePlus

Schematic diagram of the Camphor-Off switch system.
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fig2: Schematic diagram of the Camphor-Off switch system.

Mentions: We assessed the performance of the Camphor-Off system by combining the two components, cam-TA and camPr, described in the previous sections. In an effective system, we expected that a reporter gene under the control of camPr would express in the absence of camphor, a ligand of CamR, but that the expression of the reporter would be repressed by a certain amount of camphor (Figure 2). To evaluate the performance of the system, we used GFP as a reporter.


Development of a Tightly Controlled Off Switch for Saccharomyces cerevisiae Regulated by Camphor, a Low-Cost Natural Product.

Ikushima S, Zhao Y, Boeke JD - G3 (Bethesda) (2015)

Schematic diagram of the Camphor-Off switch system.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4592980&req=5

fig2: Schematic diagram of the Camphor-Off switch system.
Mentions: We assessed the performance of the Camphor-Off system by combining the two components, cam-TA and camPr, described in the previous sections. In an effective system, we expected that a reporter gene under the control of camPr would express in the absence of camphor, a ligand of CamR, but that the expression of the reporter would be repressed by a certain amount of camphor (Figure 2). To evaluate the performance of the system, we used GFP as a reporter.

Bottom Line: 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.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.

View Article: PubMed Central - PubMed

Affiliation: High Throughput Biology Center and Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205 Central Laboratories for Key Technologies, KIRIN Company Limited, Yokohama, Kanagawa 236-0004, Japan.

No MeSH data available.


Related in: MedlinePlus