Modules for cloning-free chromatin tagging in Saccharomyces cerevisae

Yeast. 2008 Mar;25(3):235-9. doi: 10.1002/yea.1580.


We describe a straightforward two-step PCR-based method to insert arrays of lac or tet operators (lacO or tetO) at specific loci in the budding yeast genome. The method entails insertion of a marker generated by PCR with classical long primers recognizing the locus of interest, followed by the replacement of this marker by a linearized plasmid bearing an array of lacI- or tetR-binding motifs. Using this technique, loci located either in the yeast genome or on yeast artificial chromosomes can be efficiently tagged. We provide a set of plasmids with different markers for cloning-free integration of lacO or tetO repeats into the yeast genome.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromatin / genetics*
  • DNA, Fungal / genetics
  • Gene Targeting
  • Genetic Engineering
  • Genetic Vectors*
  • Lac Operon
  • Operator Regions, Genetic
  • Polymerase Chain Reaction
  • Recombination, Genetic
  • Saccharomyces cerevisiae / genetics*
  • Sequence Tagged Sites*


  • Chromatin
  • DNA, Fungal