Modules for cloning-free chromatin tagging in Saccharomyces cerevisae.
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.
Rohner
Sabine
S
Friedrich Miescher Institute for Biomedical Research, Maulbeerstrasse 66, CH-4058 Basel, Switzerland.
Gasser
Susan M
SM
Meister
Peter
P
D002843:Chromatin / Q000235:genetics*; D004271:DNA, Fungal / Q000235:genetics; D018390:Gene Targeting; D005818:Genetic Engineering; D005822:Genetic Vectors; D007763:Lac Operon; D009875:Operator Regions, Genetic; D016133:Polymerase Chain Reaction; D011995:Recombination, Genetic; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D016324:Sequence Tagged Sites
D002843:Chromatin; D004271:DNA, Fungal