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ACS Synth Biol. 2015 Jul 17;4(7):853-9. doi: 10.1021/sb500372z. Epub 2015 Mar 23.

Yeast Golden Gate (yGG) for the Efficient Assembly of S. cerevisiae Transcription Units.

Author information

1
†Institute for Systems Genetics, Department of Biochemistry and Molecular Pharmacology, New York University School of Medicine, 550 First Avenue, New York, New York 10016, United States.
2
‡High Throughput Biology Center, School of Medicine, Johns Hopkins University, Edward D. Miller Research Building, 733 North Broadway, Baltimore, Maryland 21205, United States.
3
§School of Biological Sciences, University of Edinburgh, The King's Buildings, Edinburgh, EH9 3JR, United Kingdom.

Abstract

We have adapted the Golden Gate DNA assembly method to the assembly of transcription units (TUs) for the yeast Saccharomyces cerevisiae, in a method we call yeast Golden Gate (yGG). yGG allows for the easy assembly of TUs consisting of promoters (PRO), coding sequences (CDS), and terminators (TER). Carefully designed overhangs exposed by digestion with a type IIS restriction enzyme enable virtually seamless assembly of TUs that, in principle, contain all of the information necessary to express a gene of interest in yeast. We also describe a versatile set of yGG acceptor vectors to be used for TU assembly. These vectors can be used for low or high copy expression of assembled TUs or integration into carefully selected innocuous genomic loci. yGG provides synthetic biologists and yeast geneticists with an efficient new means by which to engineer S. cerevisiae.

KEYWORDS:

DNA assembly; S. cerevisiae; synthetic biology; transcription unit; yeast Golden Gate

PMID:
25756291
DOI:
10.1021/sb500372z
[Indexed for MEDLINE]

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