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Published on: May 28, 2019
Constitutive promoter modules for PCR-based gene modification in Saccharomyces cerevisiae
D J DeMarini1, E M Carlin, G P Livi
1Department of Comparative Genetics, SmithKline Beecham Pharmaceuticals, King of Prussia, PA 19406, USA. Douglas_J_Demarini@sbphrd.com
Yeast (Chichester, England)
|May 30, 2001
Summary
Researchers developed new PCR modules and plasmids for yeast gene function studies. These tools simplify gene deletion, overexpression, and localization by enabling targeted genomic modifications with minimal primers.
Area of Science:
- Molecular Biology
- Yeast Genetics
Background:
- Investigating gene function commonly involves gene deletion, overexpression, and determining subcellular localization.
- PCR modules with selectable markers and genetic elements facilitate these investigations.
Purpose of the Study:
- To describe novel plasmids for yeast gene manipulation.
- To enable efficient integration of constitutive and inducible promoters.
Main Methods:
- Development of PCR modules containing selectable markers and genetic elements.
- Design of plasmids with a truncated ADH1 promoter and N-terminal 3HA/GST tags.
- Utilizing shared vector sequences with GAL1 promoter plasmids.
Main Results:
- Creation of PCR substrates for targeted yeast genome modification.
- Plasmids allow for the integration of both constitutive (ADH1) and inducible (GAL1) promoters.
- Simplified introduction of genetic elements like epitope tags.
Conclusions:
- The developed plasmids and PCR modules streamline gene function studies in yeast.
- This approach facilitates targeted genomic locus modification.
- Enables efficient combination of different promoter types for gene expression studies.
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