Epitope tagging of yeast genes using a PCR-based strategy: More tags and improved practical routines

Epitope tagging of proteins as a strategy for the analysis of function, int eractions and the subcellular distribution of proteins has become widely us ed. In the yeast Saccharomyces cerevisiae, molecular biological techniques have been developed that use a simple PCR-based strategy to introduce epito pe tags to chromosomal loci (Wach et al., 1994). To further employ the powe r of this strategy, a variety of novel tags was constructed. These tags wer e combined with different selectable marker genes, resulting in PCR amplifi cable modules. Only one set of primers is required for the amplification of any module. Furthermore, convenient laboratory techniques are described th at facilitate the genetic manipulations of yeast strains, as well as the an alysis of the epitope-tagged proteins. Copyright (C) 1999 John Wiley & Sons , Ltd.