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

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          Abstract

          Epitope tagging of proteins as a strategy for the analysis of function, interactions and the subcellular distribution of proteins has become widely used. In the yeast Saccharomyces cerevisiae, molecular biological techniques have been developed that use a simple PCR-based strategy to introduce epitope tags to chromosomal loci (Wach et al., 1994). To further employ the power of this strategy, a variety of novel tags was constructed. These tags were combined with different selectable marker genes, resulting in PCR amplificable modules. Only one set of primers is required for the amplification of any module. Furthermore, convenient laboratory techniques are described that facilitate the genetic manipulations of yeast strains, as well as the analysis of the epitope-tagged proteins.

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          Author and article information

          Journal
          Yeast
          Yeast (Chichester, England)
          Wiley
          0749-503X
          0749-503X
          Jul 1999
          : 15
          : 10B
          Affiliations
          [1 ] The Beatson Institute for Cancer Research, CRC Beatson Laboratories, Garscube Estate, Switchback Road, Bearsden, Glasgow G61 1BD, U.K.
          Article
          10.1002/(SICI)1097-0061(199907)15:10B<963::AID-YEA399>3.0.CO;2-W
          10.1002/(SICI)1097-0061(199907)15:10B<963::AID-YEA399>3.0.CO;2-W
          10407276
          4cf5ae2f-7ccb-4bee-945f-716b46a66470
          Copyright 1999 John Wiley & Sons, Ltd.
          History

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