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The Ashbya gossypii EF‐1α promoter of the ubiquitously used MX cassettes is toxic to Saccharomyces cerevisiae
Author(s) -
Babazadeh Roja,
Moghadas Jafari Soode,
Zackrisson Martin,
Blomberg Anders,
Hohmann Stefan,
Warringer Jonas,
Krantz Marcus
Publication year - 2011
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2011.10.029
Subject(s) - saccharomyces cerevisiae , heterologous , biology , gene , promoter , microbiology and biotechnology , genetics , gene expression
Protein overexpression based on introduction of multiple gene copies is well established. To improve purification or quantification, proteins are typically fused to peptide tags. In Saccharomyces cerevisiae , this has been hampered by multicopy toxicity of the TAP and GFP cassettes used in the global strain collections. Here, we show that this effect is due to the EF‐1α promoter in the HIS3MX marker cassette rather than the tags per se . This promoter is frequently used in heterologous marker cassettes, including HIS3MX , KanMX , NatMX , PatMX and HphMX . Toxicity could be eliminated by promoter replacement or exclusion of the marker cassette. To our knowledge, this is the first report of toxicity caused by introduction of a heterologous promoter alone.