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MemStar: A one-shot Escherichia coli-based approach for high-level bacterial membrane protein production

DOI: 10.1016/j.febslet.2014.08.025 DOI Help
PMID: 25176409 PMID Help

Authors: Chiara Lee (Imperial College London) , Haejoo Kang (Imperial College London) , Anna Hjelm (Stockholm University) , Abdul Aziz Qureshi (Stockholm University) , Emmanuel Nji (Stockholm University) , Hassanul Choudhury (Diamond Light Source) , Konstantinos Beis (Diamond Light Source) , Jan-Willam De Gier (Membrane Protein Laboratory, Imperial College, London) , David Drew (Imperial College London)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Febs Letters , VOL 588 (20) , PAGES 3761 - 3769

State: Published (Approved)
Published: October 2014

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.

Journal Keywords: Membrane protein production; X-ray crystallography; High-throughput; Escherichia coli

Diamond Keywords: Bacteria

Subject Areas: Biology and Bio-materials, Technique Development

Diamond Offline Facilities: Membrane Protein Laboratory (MPL)
Technical Areas:

Added On: 01/02/2016 15:48

Discipline Tags:

Technique Development - Life Sciences & Biotech Life Sciences & Biotech

Technical Tags: