New modules for the repeated internal and N-terminal epitope tagging of genes in Saccharomyces cerevisiae.
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.
Gauss
Robert
R
Max-Delbrück Centrum für Molekulare Medizin, Robert-Rössle Str. 10, D-13092, Berlin, Germany.
Trautwein
Mark
M
Sommer
Thomas
T
Spang
Anne
A
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*
D029701:Saccharomyces cerevisiae Proteins