A flexible protein linker improves the function of epitope-tagged proteins in Saccharomyces cerevisiae

Michelle Sabourin; Creighton T. Tuzon; Timothy S. Fisher; Virginia A. Zakian

doi:10.1002/yea.1431

A flexible protein linker improves the function of epitope-tagged proteins in Saccharomyces cerevisiae

Michelle Sabourin, Creighton T. Tuzon, Timothy S. Fisher, Virginia A. Zakian

Molecular Biology

Research output: Contribution to journal › Article › peer-review

83 Scopus citations

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.

Original language	English (US)
Pages (from-to)	39-45
Number of pages	7
Journal	Yeast
Volume	24
Issue number	1
DOIs	https://doi.org/10.1002/yea.1431
State	Published - Jan 2007

All Science Journal Classification (ASJC) codes

Applied Microbiology and Biotechnology
Bioengineering
Genetics
Biochemistry
Biotechnology

Keywords

Epitope tag
In vivo function
Protein linker
Telomere
Yeast telomerase

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10.1002/yea.1431

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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.",

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AB - 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.

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KW - Yeast telomerase

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A flexible protein linker improves the function of epitope-tagged proteins in Saccharomyces cerevisiae

Abstract

All Science Journal Classification (ASJC) codes

Keywords

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