| Literature DB >> 25176409 |
Chiara Lee1, Hae Joo Kang1, Anna Hjelm2, Abdul Aziz Qureshi2, Emmanuel Nji2, Hassanul Choudhury3, Konstantinos Beis3, Jan-Willem de Gier2, David Drew4.
Abstract
Optimising membrane protein production yields in Escherichiacoli 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.Entities:
Keywords: Escherichia coli; High-throughput; Membrane protein production; X-ray crystallography
Mesh:
Substances:
Year: 2014 PMID: 25176409 DOI: 10.1016/j.febslet.2014.08.025
Source DB: PubMed Journal: FEBS Lett ISSN: 0014-5793 Impact factor: 4.124