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

Neta Agmon, Leslie A. Mitchell, Yizhi Cai, Shigehito Ikushima, James Chuang, Allen Zheng, Woo Jin Choi, J. Andrew Martin, Katrina Caravelli, Giovanni Stracquadanio, Jef D. Boeke

Research output: Contribution to journalArticle

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. (Figure Presented).

Original languageEnglish (US)
Pages (from-to)853-859
Number of pages7
JournalACS Synthetic Biology
Volume4
Issue number7
DOIs
StatePublished - Jul 17 2015

Keywords

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

ASJC Scopus subject areas

  • Biomedical Engineering
  • Biochemistry, Genetics and Molecular Biology (miscellaneous)

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  • Cite this

    Agmon, N., Mitchell, L. A., Cai, Y., Ikushima, S., Chuang, J., Zheng, A., Choi, W. J., Martin, J. A., Caravelli, K., Stracquadanio, G., & Boeke, J. D. (2015). Yeast Golden Gate (yGG) for the Efficient Assembly of S. cerevisiae Transcription Units. ACS Synthetic Biology, 4(7), 853-859. https://doi.org/10.1021/sb500372z