Saltar al contenido
Merck

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
RESUMEN

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.

MATERIALES
Referencia del producto
Marca
Descripción del producto

Sigma-Aldrich
Cloruro de sodio, for molecular biology, DNase, RNase, and protease, none detected, ≥99% (titration)
Sigma-Aldrich
Sodium chloride solution, 5 M in H2O, BioReagent, for molecular biology, suitable for cell culture
Sigma-Aldrich
Sodium chloride solution, 0.9% in water, BioXtra, suitable for cell culture
Sigma-Aldrich
Cloruro de sodio, BioReagent, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture, ≥99%
SAFC
Sodium chloride solution, 5 M
Sigma-Aldrich
Cloruro de sodio, BioXtra, ≥99.5% (AT)
Sigma-Aldrich
Sodium chloride solution, BioUltra, for molecular biology, ~5 M in H2O
Sigma-Aldrich
Cloruro de sodio, BioUltra, for molecular biology, ≥99.5% (AT)
Sigma-Aldrich
Cloruro de sodio, 99.999% trace metals basis
Sigma-Aldrich
Sodium chloride solution, 5 M
Sigma-Aldrich
Cloruro de sodio, meets analytical specification of Ph. Eur., BP, USP, 99.0-100.5%
Supelco
Cloruro de sodio, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
Cloruro de sodio, BioPerformance Certified, ≥99% (titration), suitable for insect cell culture, suitable for plant cell culture
Supelco
Cloruro de sodio, reference material for titrimetry, certified by BAM, >99.5%
Sigma-Aldrich
Cloruro de sodio, AnhydroBeads, −10 mesh, 99.999% trace metals basis
Sigma-Aldrich
Sodium chloride-35Cl, 99 atom % 35Cl
Sigma-Aldrich
Sodium chloride solution, 0.85%
Tunair shake flask, flask volume 2,500 mL, clear polypropylene, full baffle
Sigma-Aldrich
Cloruro de sodio, random crystals, optical grade, 99.9% trace metals basis
Sigma-Aldrich
Cloruro de sodio, tested according to Ph. Eur.
Sigma-Aldrich
Cloruro de sodio, tablet