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Merck

Microbial expression systems for membrane proteins.

Methods (San Diego, Calif.) (2018-04-16)
Marvin V Dilworth, Mathilde S Piel, Kim E Bettaney, Pikyee Ma, Ji Luo, David Sharples, David R Poyner, Stephane R Gross, Karine Moncoq, Peter J F Henderson, Bruno Miroux, Roslyn M Bill
摘要

Despite many high-profile successes, recombinant membrane protein production remains a technical challenge; it is still the case that many fewer membrane protein structures have been published than those of soluble proteins. However, progress is being made because empirical methods have been developed to produce the required quantity and quality of these challenging targets. This review focuses on the microbial expression systems that are a key source of recombinant prokaryotic and eukaryotic membrane proteins for structural studies. We provide an overview of the host strains, tags and promoters that, in our experience, are most likely to yield protein suitable for structural and functional characterization. We also catalogue the detergents used for solubilization and crystallization studies of these proteins. Here, we emphasize a combination of practical methods, not necessarily high-throughput, which can be implemented in any laboratory equipped for recombinant DNA technology and microbial cell culture.

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Sigma-Aldrich
麦芽糖 溶液, for molecular biology, BioReagent, ~20% in H2O
Sigma-Aldrich
吲哚-3-丁酸 钾盐, suitable for plant cell culture, BioReagent
Sigma-Aldrich
四乙二醇单辛醚, liquid, ≥98% (GC)
Sigma-Aldrich
五聚乙二醇单癸醚, BioXtra, ≥97.0% (GC)