Journal
YEAST
Volume 25, Issue 3, Pages 235-239Publisher
JOHN WILEY & SONS LTD
DOI: 10.1002/yea.1580
Keywords
nucleus; chromatin; tagging; lacO; tetO; CFC tags
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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 (c) 2008 John Wiley & Sons, Ltd.
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