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Curing of the 2 mu DNA plasmid from Saccharomyces cerevisiae
Journal of Bacteriology
|March 1, 1981
Abstract:
The 2 mu DNA plasmid is often eliminated from yeast cells when they are transformed with the 2 mu DNA-LEU2-pMB9 composite plasmid pJDB219. Since pJDB219 is subsequently lost with high frequency, derivatives lacking all 2 mu DNA can be prepared from any strain.
Insights
The 2-micrometer (µm) DNA plasmid is frequently lost from yeast cells after transformation with the pJDB219 composite plasmid. This high loss rate allows for the creation of yeast strains lacking any 2-micrometer DNA.
Area of Science:
- Molecular Biology
- Yeast Genetics
Background:
- The 2-micrometer (µm) DNA is a high-copy-number yeast plasmid essential for its own replication and maintenance.
- Transformation of yeast with composite plasmids like pJDB219, which contains 2 µm DNA and other elements, can lead to instability.
Purpose of the Study:
- To investigate the stability and segregation of the 2 µm DNA component within the pJDB219 composite plasmid in yeast.
- To explore methods for generating yeast strains devoid of endogenous 2 µm DNA.
Main Methods:
- Yeast transformation with the pJDB219 plasmid.
- Monitoring plasmid retention and loss over yeast generations.
- Characterization of yeast derivatives lacking 2 µm DNA.
Main Results:
- The 2 µm DNA component of pJDB219 is frequently eliminated from transformed yeast cells.
- The composite plasmid pJDB219 exhibits high-frequency loss during yeast cultivation.
- Yeast strains lacking all 2 µm DNA can be successfully derived from pJDB219 transformants.
Conclusions:
- The instability of pJDB219 facilitates the generation of 2 µm DNA-free yeast strains.
- This process offers a practical approach for obtaining yeast derivatives without endogenous 2 µm DNA for research purposes.