MemStar: A one-shot Escherichia coli-based approach for high-level bacterial membrane protein production
2014 (English)In: FEBS Letters, ISSN 0014-5793, E-ISSN 1873-3468, Vol. 588, no 20, 3761-3769 p.Article in journal (Refereed) Published
Optimising membrane protein production yields in Escherichia coli can be time- and resource-consuming. Here, we present a simple and effective Membrane protein Single shot amplification recipe: MemStar. This one-shot amplification recipe is based on the E. coli strain Lemo21(DE3), the PASM-5052 auto-induction medium and, contradictorily, an IPTG induction step. Using MemStar, production yields for most bacterial membrane proteins tested were improved to reach an average of 5 mg L-1 per OD600 unit, which is significantly higher than yields obtained with other common production strategies. With MemStar, we have been able to obtain new structural information for several transporters, including the sodium/proton antiporter NapA. (C) 2014 Federation of European Biochemical Societies.
Place, publisher, year, edition, pages
2014. Vol. 588, no 20, 3761-3769 p.
Membrane protein production, X-ray crystallography, High-throughput, Escherichia coli
Biochemistry and Molecular Biology Cell Biology
Research subject Biochemistry
IdentifiersURN: urn:nbn:se:su:diva-108362DOI: 10.1016/j.febslet.2014.08.025ISI: 000341993000015OAI: oai:DiVA.org:su-108362DiVA: diva2:758198