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Related Experiment Videos

Yeast vectors for integration at the HO locus.

W P Voth1, J D Richards, J M Shaw

  • 1Department of Pathology and Department of Biology, University of Utah, Salt Lake City, UT 84132, USA.

Nucleic Acids Research
|June 19, 2001
PubMed
Summary

New yeast vectors enable targeted DNA integration at the Saccharomyces cerevisiae HO locus for neutral genetic modifications. These vectors utilize KanMX or hisG-URA3-hisG markers for selection, facilitating precise genetic engineering in yeast and bacteria.

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Area of Science:

  • Molecular Biology
  • Yeast Genetics
  • Genetic Engineering

Background:

  • Targeted DNA integration is crucial for genetic engineering.
  • The Saccharomyces cerevisiae HO locus is a site for neutral integration.
  • Efficient selection markers are needed for identifying successful integrants.

Purpose of the Study:

  • To develop novel yeast vectors for targeted integration at the HO locus.
  • To create vectors with selectable markers for yeast and bacterial systems.
  • To facilitate neutral genetic modifications in Saccharomyces cerevisiae.

Main Methods:

  • Construction of yeast vectors with KanMX and hisG-URA3-hisG selectable markers.
  • Transformation of Saccharomyces cerevisiae and selection of integrants.

Related Experiment Videos

  • Use of G418 for KanMX selection and uracil prototrophy/auxotrophy for hisG-URA3-hisG selection.
  • Development of bacterial polylinker vectors derived from pUC21 and pUK21.

    • Main Results:

    • Successfully constructed yeast vectors for targeted integration at the HO locus.
    • Demonstrated neutral integration with no adverse effects on yeast growth.
    • Established selection protocols using G418 resistance and uracil auxotrophy.
    • Developed new bacterial polylinker vectors with ampicillin and kanamycin resistance.

    Conclusions:

    • The new yeast vectors provide a robust system for neutral, targeted DNA integration in Saccharomyces cerevisiae.
    • The developed vectors and selection strategies simplify genetic manipulation in yeast.
    • The new bacterial vectors expand options for cloning and genetic engineering in bacterial systems.