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Yeast-gene replacement using PCR products.
Megan Bergkessel1, Christine Guthrie, John Abelson
1Department of Biochemistry and Biophysics, University of California, San Francisco, CA, USA.
Methods in Enzymology
|November 5, 2013
Summary
Yeast gene replacement using selectable markers facilitates gene knockout studies and genomic region analysis. This method aids in assessing gene function loss and modifying noncoding DNA elements.
Area of Science:
- Molecular Biology
- Yeast Genetics
Background:
- Gene replacement in yeast is crucial for understanding gene function.
- Selectable markers simplify the identification of modified yeast strains.
Purpose of the Study:
- To describe a method for yeast gene replacement using selectable markers.
- To enable functional assessment of gene knockouts.
- To facilitate modifications of noncoding genomic regions.
Main Methods:
- Replacing a genomic region with a selectable marker.
- Assessing gene function through knockout.
- Modifying promoters and untranslated regions (UTRs).
Main Results:
- Successful identification of yeast cells with gene replacement.
- Enables assessment of functional consequences of gene deletion.
- Allows for targeted modification of noncoding DNA.
Conclusions:
- Selectable marker-based gene replacement is an effective technique in yeast.
- This method supports diverse genomic manipulation strategies.
- It is valuable for functional genomics and genetic engineering in Saccharomyces cerevisiae.
Keywords:
Agarose gel visualizationColonies growing on selective mediaDeoxyribonucleic acid (DNA)Polyacrylamide gel electrophoresis (PAGE)Polymerase chain reaction (PCR)Transforming yeast with linear PCR productYeast-gene replacement
