Journal
ACS SYNTHETIC BIOLOGY
Volume 4, Issue 7, Pages 853-859Publisher
AMER CHEMICAL SOC
DOI: 10.1021/sb500372z
Keywords
synthetic biology; DNA assembly; S. cerevisiae; transcription unit; yeast Golden Gate
Categories
Funding
- National Science Foundation grant [MCB-0718846]
- Defense Advanced Research Projects Agency [N66001-12-C-4020]
- Natural Sciences and Engineering Research Council of Canada
- BBSRC [BB/M005690/1] Funding Source: UKRI
- Biotechnology and Biological Sciences Research Council [BB/M005690/1] Funding Source: researchfish
- Direct For Biological Sciences
- Div Of Molecular and Cellular Bioscience [1443299, 1445537] Funding Source: National Science Foundation
Ask authors/readers for more resources
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.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available