Neochromosomes: Creating transcriptional units with yeast Golden Gate assembly
Author(s): Eric Cooper
Hartwick College
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- Workflow for yeast Golden Gate assembly.docx(DOCX | 680 KB)
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Description
This resource is a series of protocols using the yeast Golden Gate (yGG) method as developed by Neta Agmon and Leslie Mitchell in Jef Boeke’s lab to assemble a functional yeast gene with protein-coding and regulatory sequences. First, there are protocols to assemble a single gene whose activity is easy to measure (e.g. an auxotrophic marker, antibiotic resistance gene, LacZ, etc.) just to get the system up and running in your lab. Once you do, you can apply the methodology to any gene(s) of interest (i.e. it does not have to be a yeast gene). You can also use a variation of this method to string together multiple genes (via VEGAS assembly) into a single plasmid to study metabolic pathways. This resource was developed as part of NSF Award #1827204, RCN-UBE: The Build-a-Genome Network.
Cite this work
Researchers should cite this work as follows:
- Eric Cooper (2019). Neochromosomes: Creating transcriptional units with yeast Golden Gate assembly. Build A Genome Network, (Version 2.0). QUBES Educational Resources. doi:10.25334/3J95-S075