title: PCR-mediated epitope tagging of genes in yeast.
abstract: Epitope tagging of genes is a powerful technique facilitating assays for gene function, determination of subcellular distribution of proteins, affinity purification, study of protein interaction with other proteins, DNA or RNA, and any other antibody-based approach in the absence of protein-specific antibodies. Here, we describe a one-step PCR-based strategy for insertion of epitope tags at the chromosomal locus. This method takes advantage of efficient homologous recombination in yeast. PCR amplified tags are directed to desired chromosomal loci with the help of primer-encoded flanking homologous sequences enabling selective epitope tagging of genes of interest.
authors.affiliation: Department of Biological Chemistry, School of Medicine, University of California Irvine, 240D Med Sci I, Irvine, CA, 92697-1700, USA.
authors.forename: Radhika
authors.initials: R
authors.lastname: Mathur
authors.forename: Peter
authors.initials: P
authors.lastname: Kaiser
mesh_terms: D015153:Blotting, Western / Q000379:methods; D015825:Chromosomes, Fungal; D017931:DNA Primers / Q000235:genetics; D018604:Epitope Mapping; D000939:Epitopes / Q000235:genetics*; D005656:Fungal Proteins / Q000235:genetics*; D005800:Genes, Fungal; D056426:Genetic Loci; D059765:Homologous Recombination; D016133:Polymerase Chain Reaction / Q000379:methods*; D014170:Transformation, Genetic; D015003:Yeasts / Q000235:genetics*
chemical_list: D017931:DNA Primers; D000939:Epitopes; D005656:Fungal Proteins