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Blount BA, et al.  (2012) Rational diversification of a promoter providing fine-tuned expression and orthogonal regulation for synthetic biology. PLoS One 7(3):e33279

Abstract: Yeast is an ideal organism for the development and application of synthetic biology, yet there remain relatively few well-characterised biological parts suitable for precise engineering of this chassis. In order to address this current need, we present here a strategy that takes a single biological part, a promoter, and re-engineers it to produce a fine-graded output range promoter library and new regulated promoters desirable for orthogonal synthetic biology applications. A highly constitutive Saccharomyces cerevisiae promoter, PFY1p, was identified by bioinformatic approaches, characterised in vivo and diversified at its core sequence to create a 36-member promoter library. TetR regulation was introduced into PFY1p to create a synthetic inducible promoter (iPFY1p) that functions in an inverter device. Orthogonal and scalable regulation of synthetic promoters was then demonstrated for the first time using customisable Transcription Activator-Like Effectors (TALEs) modified and designed to act as orthogonal repressors for specific PFY1-based promoters. The ability to diversify a promoter at its core sequences and then independently target Transcription Activator-Like Orthogonal Repressors (TALORs) to virtually any of these sequences shows great promise toward the design and construction of future synthetic gene networks that encode complex "multi-wire" logic functions.

Status: Published Type: Journal Article PubMed ID: 22442681

Topics addressed in this paper

Number of different genes curated to this paper: 17

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Topics Topics not linked to Genes Genes linked to topics (#1 - 10 )
ADH1 ALA1 ARA1 BIO2 CHO1 CIT2 CYC1 DBP5 GIM5 HRT1
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Topics Genes linked to topics (#11 - 17 )
MDH3 PFY1 RPL6A RPN6 VMA6 YKT6 YSA1
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