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Microbial expression systems for membrane proteins
Methods ( IF 4.8 ) Pub Date : 2018-09-01 , DOI: 10.1016/j.ymeth.2018.04.009 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
Methods ( IF 4.8 ) Pub Date : 2018-09-01 , DOI: 10.1016/j.ymeth.2018.04.009 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
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.
中文翻译:
膜蛋白微生物表达系统
尽管取得了许多引人注目的成功,但重组膜蛋白的生产仍然是一项技术挑战。与可溶性蛋白质相比,已发表的膜蛋白结构仍然少得多。然而,由于已经开发出经验方法来产生这些具有挑战性的目标所需的数量和质量,因此正在取得进展。本综述重点关注微生物表达系统,该系统是用于结构研究的重组原核和真核膜蛋白的关键来源。我们概述了宿主菌株、标签和启动子,根据我们的经验,它们最有可能产生适合结构和功能表征的蛋白质。我们还对用于这些蛋白质的溶解和结晶研究的去污剂进行了分类。这里,
更新日期:2018-09-01
中文翻译:
膜蛋白微生物表达系统
尽管取得了许多引人注目的成功,但重组膜蛋白的生产仍然是一项技术挑战。与可溶性蛋白质相比,已发表的膜蛋白结构仍然少得多。然而,由于已经开发出经验方法来产生这些具有挑战性的目标所需的数量和质量,因此正在取得进展。本综述重点关注微生物表达系统,该系统是用于结构研究的重组原核和真核膜蛋白的关键来源。我们概述了宿主菌株、标签和启动子,根据我们的经验,它们最有可能产生适合结构和功能表征的蛋白质。我们还对用于这些蛋白质的溶解和结晶研究的去污剂进行了分类。这里,