Curing Saccharomyces cerevisiae of the 2 micron plasmid by targeted DNA damage

Ephraim L. Tsalik, Marc R. Gartenberg

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41 Scopus citations


Powerful mutagenic screens of yeast Saccharomyces cerevisiae have recently been developed which require strains that lack the endogenous 2 micron plasmid (Burns et al., 1994). Here, we describe a simple and reliable method for curing yeast of the highly stable genetic element. The approach employs heterologous expression of a 'step-arrest' mutant of the F1p recombinase. The mutant, F1p H305L (Parsons et al., 1988), forms long-lived covalent protein-DNA complexes exclusively at 2 micron-borne recombinase target sites. In vivo, the complexes serve as sites of targeted DNA damage. Using Southern hybridization and a colony color assay for plasmid loss, we show that expression of the mutant enzyme results in the effective elimination of the 2 micron from cells.

Original languageEnglish (US)
Pages (from-to)847-852
Number of pages6
Issue number9
StatePublished - Jun 30 1998

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Genetics


  • 2 micron plasmid
  • Curing
  • DNA damage
  • F1p
  • Saccharomyces cerevisiae


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