Current Genetics

, Volume 27, Issue 4, pp 306–308

Strategy for deletion of complete open reading frames in Saccharomyces cerevisiae

  • Ines Eberhardt
  • Stefan Hohmann
Original Paper

DOI: 10.1007/BF00352097

Cite this article as:
Eberhardt, I. & Hohmann, S. Curr Genet (1995) 27: 306. doi:10.1007/BF00352097

Abstract

The classical disruption method for yeast genes is by using in vitro deletion of the gene of interest, or of a part of it, with restriction enzymes. We are now routinely using a strategy that takes advantage of polymerase chain reactions (PCRs) which amplify large pieces of DNA. Since this approach results in a complete, precise deletion of the open reading frame, which is replaced by a unique restriction site, the ligated PCR can be used for the insertion of different markers of for two-step gene disruptions without an inserted marker. As we have now used this strategy for the deletion of more than ten genes we have in this report included some hints based on our experience.

Key words

Gene deletion Open reading frame Saccharomyces cerevisiae Polymerase chain reaction 

Copyright information

© Springer-Verlag 1995

Authors and Affiliations

  • Ines Eberhardt
    • 1
  • Stefan Hohmann
    • 1
  1. 1.Laboratorium voor Moleculaire CelbiologieKatholieke Universiteit te LeuvenLeuven-Heverlee, FlandersBelgium

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