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A flexible protein linker improves the function of epitope‐tagged proteins in Saccharomyces cerevisiae
Author(s) -
Sabourin Michelle,
Tuzon Creighton T.,
Fisher Timothy S.,
Zakian Virginia A.
Publication year - 2007
Publication title -
yeast
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 102
eISSN - 1097-0061
pISSN - 0749-503X
DOI - 10.1002/yea.1431
Subject(s) - epitope , schizosaccharomyces pombe , saccharomyces cerevisiae , open reading frame , biology , linker , computational biology , function (biology) , amino acid , gene , biochemistry , genetics , peptide sequence , antibody , computer science , operating system
Abstract 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 . Copyright © 2007 John Wiley & Sons, Ltd.