- MemStar: a one-shot Escherichia coli-based approach for high-level bacterial membrane protein production.
MemStar: a one-shot Escherichia coli-based approach for high-level bacterial membrane protein production.
FEBS letters (2014-09-02)
Chiara Lee, Hae Joo Kang, Anna Hjelm, Abdul Aziz Qureshi, Emmanuel Nji, Hassanul Choudhury, Konstantinos Beis, Jan-Willem de Gier, David Drew
PMID25176409
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.
MATERIALS
Product Number
Brand
Product Description
Sigma-Aldrich
Sodium chloride, BioReagent, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture, ≥99%
Sigma-Aldrich
Sodium chloride, for molecular biology, DNase, RNase, and protease, none detected, ≥99% (titration)
Tunair™ shake flask, flask volume 2,500 mL, clear polypropylene, full baffle
Sigma-Aldrich
Sodium chloride, BioPerformance Certified, ≥99% (titration), suitable for insect cell culture, suitable for plant cell culture
Sigma-Aldrich
Sodium chloride solution, 5 M in H2O, BioReagent, for molecular biology, suitable for cell culture