A flexible protein linker improves the function of epitope-tagged proteins in Saccharomyces cerevisiae.
Epitope tagging permits the detection of proteins when protein-specific antibodies are not available. However, the epitope tag can reduce the function of the tagged protein. Here we describe a cassette that can be used to introduce an eight amino acid flexible linker between multiple Myc epitopes and the open reading frame of a given gene. We show that inserting the linker improves the in vivo ability of the telomerase subunits Est2p and Est1p to maintain telomere length. The methods used here are generally applicable to improve the function of tagged proteins in both Saccharomyces cerevisiae and Schizosaccharomyces pombe.
Sabourin
Michelle
M
Department of Molecular Biology, Princeton University, Princeton, NJ 08544, USA.
Tuzon
Creighton T
CT
Fisher
Timothy S
TS
Zakian
Virginia A
VA
D015153:Blotting, Western; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics; D000939:Epitopes / Q000235:genetics / Q000378:metabolism*; D016259:Genes, myc; D016133:Polymerase Chain Reaction; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D019098:Telomerase / Q000235:genetics / Q000378:metabolism*; D016615:Telomere / Q000502:physiology
D004271:DNA, Fungal; D000939:Epitopes; D029701:Saccharomyces cerevisiae Proteins; D019098:Telomerase