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Construction of a yeast plasmid cloning vector with high stability inSaccharomyces cerevisiae strains deficient in 2 µmDNA

Konstruktion eines Hefe-Plasmid-Cloningvektors mit hoher Stabilität in 2 µm DNA-defizientenSaccharomyces cerevisiae-Stämmen

  • Organische Chemie Und Biochemie
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Abstract

PlasmidpCP1 was constructed from cloned 2 µmDNA ofSaccharomyces cerevisiae and from plasmidpJDB207. VectorpCP1 contains the completeB form of 2 µmDNA interrupted in theFLP gene, together withDNA derived from theEscherichia coli plasmidpAT153 and a low expression variant of theS. cerevisiae LEU2 gene. The new vector is lost at a low frequency from yeastcir + orcir 0 strains under non-selective growth conditions and is stable against rearrangements incir 0 strains. Its usefulness for curingcir + strains from endogenous 2 µmDNA and for their conversion tocir 0 strains was demonstrated.

Zusammenfassung

PlasmidpCP1 wurde ausgehend von klonierterSaccharomyces cerevisiae-2 µmDNA und PlasmidpJDB207 konstruiert. Der Vektor enthält die vollständige, imFLP-Gen unterbrocheneB-Form der 2 µmDNA, vomEscherichia coli-Plasmidp AT153 abgeleiteteDNA und eine wenig aktive Variante desS. cerevisiae-LEU2-Gens. Der neue Vektor weist unter nichtselektiven Wachstumsbedingungen incir +- undcir 0-Stämmen eine niedrige Verlustrate auf und ist incir 0-Stämmen stabil gegen Umlagerungen. Seine Verwendbarkeit für die Umwandlung voncir +-Stämmen incir 0-Stämme durch Verdrängung endogener 2 µm-DNA wurde nachgewiesen.

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Authors and Affiliations

  1. Institut für Allgemeine Biochemie der Universität Wien and Ludwig-Boltzmann-Forschungsstelle für Biochemie, Wien, A-1090, Vienna, Austria

    Andreas Bachmair & Helmut Ruis

Authors
  1. Andreas Bachmair
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  2. Helmut Ruis
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Dedicated to Professor Dr.Karl Schlögl on the occasion of his 60th birthday.

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Bachmair, A., Ruis, H. Construction of a yeast plasmid cloning vector with high stability inSaccharomyces cerevisiae strains deficient in 2 µmDNA . Monatsh Chem 115, 1229–1235 (1984). https://doi.org/10.1007/BF00809354

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  • DOI: https://doi.org/10.1007/BF00809354

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