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Related Experiment Videos

Multifunctional yeast high-copy-number shuttle vectors.

T W Christianson1, R S Sikorski, M Dante

  • 1Department of Microbiology, Southern Illinois University, Carbondale 62901.

Gene
|January 2, 1992
PubMed
Summary
This summary is machine-generated.

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Researchers developed new high-copy-number yeast episomal plasmid (YEp) vectors, the pRS420 series. These vectors offer selectable markers for Saccharomyces cerevisiae and are based on a versatile phagemid backbone for E. coli manipulation.

Area of Science:

  • Molecular Biology
  • Yeast Genetics
  • Plasmid Technology

Background:

  • Development of novel cloning and expression vectors is crucial for yeast research.
  • Existing yeast episomal plasmid (YEp) vectors have limitations in copy number or backbone utility.
  • The Saccharomyces cerevisiae 2-micron (2-micron) plasmid is a key element for high-copy-number episomal vectors in yeast.

Purpose of the Study:

  • To construct and characterize a new series of high-copy-number yeast shuttle vectors.
  • To provide YEp vectors with diverse yeast selectable markers (HIS3, TRP1, LEU2, URA3).
  • To integrate the utility of a multifunctional phagemid backbone for molecular manipulation.

Main Methods:

  • Construction of yeast shuttle vectors (pRS420 series) incorporating 2-micron plasmid sequences.

Related Experiment Videos

  • Utilizing the pBluescript II SK+ phagemid backbone for E. coli compatibility and in vitro manipulation.
  • Assessing plasmid copy number and mitotic segregation rates in Saccharomyces cerevisiae.

    • Main Results:

    • Successfully constructed the pRS420 series of YEp vectors with HIS3, TRP1, LEU2, or URA3 selectable markers.
    • The pRS420 plasmids exhibit a high copy number (approx. 20 per cell) in yeast.
    • Mitotic segregation rates during non-selective growth are comparable to existing YEp and YCp vectors (1.5-5% loss per doubling).

    Conclusions:

    • The pRS420 series offers valuable high-copy-number YEp vectors for Saccharomyces cerevisiae.
    • These vectors combine the benefits of the 2-micron origin of replication with the versatile pBluescript II SK+ backbone.
    • The pRS420 series serves as high-copy-number counterparts to existing pRS vectors, expanding molecular biology toolkits.