Yeast Golden Gate (yGG) for the Efficient Assembly of S. cerevisiae Transcription Units
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- 作者：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
- 刊名：ACS Synthetic Biology
- 出版年：2015
- 出版时间：July 17, 2015
- 年：2015
- 卷：4
- 期：7
- 页码：853-859
- 全文大小：322K
- ISSN：2161-5063
文摘
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.