title: New modules for the repeated internal and N-terminal epitope tagging of genes in Saccharomyces cerevisiae.
abstract: Epitope tagging is a powerful method for the rapid analysis of protein function. In Saccharomyces cerevisiae epitope tags are introduced easily into chromosomal loci by homologous recombination using a simple PCR-based strategy. Although quite a number of tools exist for C-terminal tagging as well as N-terminal tagging of proteins expressed by heterologous promoters, there are only very limited possibilities to tag proteins at the N-terminus and retain the endogenous expression level. Furthermore, no PCR-templates for internal tagging have been reported. Here we describe new modules that are suitable for both the repeated N-terminal and internal tagging of proteins, leaving their endogenous promoters intact. The tags include 6xHA, 9xMyc, yEGFP, TEV-GST-6xHIS, ProtA, TEV-ProtA and TEV-ProtA-7xHIS in conjunction with different heterologous selection markers.
authors.affiliation: Max-Delbrück Centrum für Molekulare Medizin, Robert-Rössle Str. 10, D-13092, Berlin, Germany.
authors.forename: Robert
authors.initials: R
authors.lastname: Gauss
authors.forename: Mark
authors.initials: M
authors.lastname: Trautwein
authors.forename: Thomas
authors.initials: T
authors.lastname: Sommer
authors.forename: Anne
authors.initials: A
authors.lastname: Spang
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D015825:Chromosomes, Fungal; D018604:Epitope Mapping; D005800:Genes, Fungal; D008969:Molecular Sequence Data; D010957:Plasmids; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins / Q000276:immunology / Q000502:physiology*
chemical_list: D029701:Saccharomyces cerevisiae Proteins