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Detection of a 2 mu derivative yeast plasmid with altered properties
Abstract:
The Saccharomyces cerevisiae (Sacch. cerevisiae) strain RXII, like many others, harbours plasmid DNAs. and one category of them is homologous to the 2 mu plasmid of yeast. DNA-DNA hybridization experiments indicated altered structures of this species as regards the number and distribution of the restriction sites. The efforts made to clone either the whole plasmid in pBR328 or its fragments in pBR322 vectors remained unsuccessful, since deleted plasmids were isolated without insert DNA, and even the loss of vector sequences was observed. The data suggest, that the 2 mu derivative plasmid in strain RXII represent an unique category of this extrachromosomal genetic element.
Insights
This study investigates a unique Saccharomyces cerevisiae plasmid derivative homologous to the 2-micron plasmid. Cloning attempts failed, suggesting an unusual structure for this extrachromosomal genetic element.
Area of Science:
- Molecular Biology
- Yeast Genetics
- Plasmid Biology
Background:
- Saccharomyces cerevisiae (Sacch. cerevisiae) strains commonly contain plasmid DNAs.
- A subset of these plasmids are homologous to the well-characterized 2-micron plasmid of yeast.
Purpose of the Study:
- To characterize a novel 2-micron plasmid derivative found in Sacch. cerevisiae strain RXII.
- To investigate the structural and functional properties of this unique extrachromosomal genetic element.
Main Methods:
- DNA-DNA hybridization to analyze plasmid structure.
- Restriction site mapping to identify alterations.
- Plasmid cloning attempts using pBR328 and pBR322 vectors.
Main Results:
- DNA-DNA hybridization revealed altered restriction site number and distribution compared to the standard 2-micron plasmid.
- Cloning efforts were unsuccessful, yielding deleted plasmids or loss of vector sequences.
- These findings indicate significant structural deviations in the RXII strain's plasmid.
Conclusions:
- The 2-micron plasmid derivative in Sacch. cerevisiae strain RXII possesses unique structural characteristics.
- This extrachromosomal genetic element represents a distinct category of yeast plasmids.
- Further research is warranted to fully elucidate the nature of this unique plasmid.