Lee Chiara, Kang Hae Joo, Hjelm Anna, Qureshi Abdul Aziz, Nji Emmanuel, Choudhury Hassanul, Beis Konstantinos, de Gier Jan-Willem, Drew David
Division of Molecular Biosciences, Imperial College London, London SW7 2AZ, UK.
Centre for Biomembrane Research, Department of Biochemistry and Biophysics, Stockholm University, SE-106 91 Stockholm, Sweden.
FEBS Lett. 2014 Oct 16;588(20):3761-9. doi: 10.1016/j.febslet.2014.08.025. Epub 2014 Aug 28.
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.
优化大肠杆菌中膜蛋白的产量既耗时又耗资源。在此，我们提出了一种简单有效的膜蛋白单次扩增方法：MemStar。这种单次扩增方法基于大肠杆菌Lemo21(DE3)菌株、PASM - 5052自诱导培养基，并且矛盾的是，还包括一个IPTG诱导步骤。使用MemStar，大多数测试的细菌膜蛋白的产量提高到平均每OD600单位5 mg L(-1)，这显著高于其他常见生产策略所获得的产量。通过MemStar，我们已经能够获得几种转运蛋白的新结构信息，包括钠/质子反向转运蛋白NapA。
