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Replicating plasmids in Schizosaccharomyces pombe: improvement of symmetric segregation by a new genetic element

W D Heyer, M Sipiczki, J Kohli

    Molecular and Cellular Biology
    |January 1, 1986
    PubMed

    Insights

    Researchers evaluated yeast-Escherichia coli shuttle vectors in Schizosaccharomyces pombe. A stabilizing fragment (stb) was identified for the pFL20 plasmid, improving its mitotic and meiotic stability in fission yeast.

    Area of Science:

    • Molecular Biology
    • Genetics
    • Yeast Research

    Background:

    • Widely used yeast-Escherichia coli shuttle vectors are crucial for molecular biology applications.
    • Understanding vector behavior in different yeast species is essential for genetic manipulation.
    • Schizosaccharomyces pombe is a key model organism in biological research.

    Purpose of the Study:

    • To characterize the behavior of common yeast-Escherichia coli shuttle vectors in Schizosaccharomyces pombe.
    • To investigate the factors contributing to plasmid stability in S. pombe.
    • To identify elements that enhance plasmid maintenance in fission yeast.

    Main Methods:

    • Transformation of S. pombe with various shuttle vectors (pDB248, YEp13, pLEU2, pURA3, pFL20).
    • Assessment of transformation frequency, mitotic and meiotic stability, and copy number.
    • Pedigree analysis of transformed cells to measure plasmid partitioning.
    • Identification and characterization of a stabilizing DNA fragment (stb) from S. pombe.

    Main Results:

    • 2-micron vectors (pDB248, YEp13) exhibited behavior similar to Saccharomyces cerevisiae, with intermediate mitotic and low meiotic stability.
    • S. cerevisiae integration vectors (pLEU2, pURA3) replicated autonomously in S. pombe at high copy numbers, suggesting a lack of sequence specificity.
    • The pFL20 vector showed high stability, attributed to an identified S. pombe DNA fragment (stb) that improved plasmid partitioning during mitosis and meiosis.

    Conclusions:

    • The behavior of common yeast vectors varies in S. pombe compared to S. cerevisiae.
    • Autonomous replication of S. cerevisiae integration vectors in S. pombe highlights species-specific replication mechanisms.
    • The identified S. pombe stb fragment is a key element for enhancing plasmid stability in fission yeast, with potential applications in genetic engineering.

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