The Ashbya gossypiiEF-1α promoter of the ubiquitously used MX cassettes is toxic to Saccharomyces cerevisiae.
Artikel i vetenskaplig tidskrift, 2011

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.

Histidine toxicity Protein overexpression HIS3 MX toxicity TAP-tag toxicity

Författare

Roja Babazadeh

Göteborgs universitet

Soode Moghadas Jafari

Martin Zackrisson

Göteborgs universitet

Anders Blomberg

Göteborgs universitet

Stefan Hohmann

Göteborgs universitet

Jonas Warringer

Göteborgs universitet

Marcus Krantz

Göteborgs universitet

FEBS letters

1873-3468 (ISSN)

Ämneskategorier

Cellbiologi

Biokemi och molekylärbiologi

DOI

10.1016/j.febslet.2011.10.029

PubMed

22033143