1. MemStar: A one-shotEscherichia coli-based approach for high-level bacterial membrane protein production
- Author
-
Chiara Lee, Konstantinos Beis, Hassanul G. Choudhury, David A. Drew, Anna Hjelm, Emmanuel Nji, Abdul Aziz Qureshi, Jan-Willem de Gier, and Hae Joo Kang
- Subjects
Antiporter ,Sodium ,Biophysics ,chemistry.chemical_element ,lac operon ,High-throughput ,Chemical Fractionation ,Biology ,Cell Fractionation ,medicine.disease_cause ,Biochemistry ,Membrane protein production ,Industrial Microbiology ,Structural Biology ,Escherichia coli ,Genetics ,medicine ,Molecular Biology ,X-ray crystallography ,NAPA ,One shot ,Escherichia coli Proteins ,Single shot ,Cell Biology ,Membrane protein ,chemistry ,Bacterial Outer Membrane Proteins - Abstract
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 5mgL−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.
- Published
- 2014
- Full Text
- View/download PDF