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Modules for cloning‐free chromatin tagging in Saccharomyces cerevisae
Author(s) -
Rohner Sabine,
Gasser Susan M.,
Meister Peter
Publication year - 2008
Publication title -
yeast
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 102
eISSN - 1097-0061
pISSN - 0749-503X
DOI - 10.1002/yea.1580
Subject(s) - biology , genetics , plasmid , saccharomyces cerevisiae , locus (genetics) , cloning (programming) , tetr , genome , yeast , budding yeast , chromatin , computational biology , dna , gene , repressor , computer science , transcription factor , programming language
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. Copyright © 2008 John Wiley & Sons, Ltd.