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Related Experiment Videos

Copy number control by a yeast centromere.

G Tschumper, J Carbon

    Gene
    |August 1, 1983
    PubMed
    Summary
    This summary is machine-generated.

    Yeast centromere (CEN3) plasmids are maintained at low copy numbers, mimicking chromosomes. The 2-micron plasmid

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    Proceedings of the National Academy of Sciences of the United States of America·1999

    Area of Science:

    • Molecular Biology
    • Yeast Genetics
    • Plasmids

    Background:

    • Plasmids are extrachromosomal DNA elements.
    • Yeast (Saccharomyces cerevisiae) centromeres regulate chromosome segregation.
    • The yeast 2-micron plasmid is a high-copy-number element.

    Purpose of the Study:

    • To investigate the maintenance and stability of composite plasmids containing a yeast centromere (CEN3) and the yeast 2-micron plasmid.
    • To determine the copy number control mechanisms of such composite plasmids.
    • To understand the role of the 2-micron plasmid's recombination system in plasmid stability.

    Main Methods:

    • Construction of composite plasmids integrating CEN3 into the yeast 2-micron plasmid.
    • Yeast transformation and selection for plasmid maintenance.

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  • Analysis of plasmid copy number and segregation patterns during mitosis and meiosis.
  • Investigation of plasmid deletions and the role of the 2-micron plasmid's FLP recombination system.

    • Main Results:

    • Composite CEN3-2-micron plasmids are maintained at a low copy number (approximately one per haploid genome), similar to chromosomes.
    • The CEN3 copy number control is dominant over the 2-micron plasmid's amplification system.
    • These plasmids exhibit stable maintenance and Mendelian segregation during meiosis.
    • Deletions within the composite plasmids, leading to loss of CEN3 control and high copy number reversion, require the 2-micron plasmid's recombination system (FLP).

    Conclusions:

    • Yeast centromeres effectively confer low copy number maintenance and stable segregation to plasmids.
    • The yeast 2-micron plasmid's recombination system can lead to deletions in composite plasmids, overriding centromeric control.
    • These findings have implications for the design of stable yeast-based vectors for biotechnological applications.