Efficient PCR-based gene disruption in Saccharomyces strains using intergenic primers

Yeast. 2002 Mar 15;19(4):319-28. doi: 10.1002/yea.817.


Gene disruptions are a vital tool for understanding Saccharomyces cerevisiae gene function. An arrayed library of gene disruption strains has been produced by a consortium of yeast laboratories; however their use is limited to a single genetic background. Since the yeast research community works with several different strain backgrounds, disruption libraries in other common laboratory strains are desirable. We have developed simple PCR-based methods that allow transfer of gene disruptions from the S288C-derived strain library into any Saccharomyces strain. One method transfers the unique sequence tags that flank each of the disrupted genes and replaces the kanamycin resistance marker with a recyclable URA3 gene from Kluyveromyces lactis. All gene-specific PCR amplifications for this method are performed using a pre-existing set of primers that are commercially available. We have also extended this PCR technique to develop a second general gene disruption method suitable for any transformable strain of Saccharomyces.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • DNA Primers
  • Gene Deletion
  • Kluyveromyces / genetics
  • Molecular Biology
  • Polymerase Chain Reaction / methods*
  • Saccharomyces cerevisiae / genetics*
  • Transformation, Genetic


  • DNA Primers