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值得关注的技术:结构生物学中蛋白质生产的替代宿主。

Technologies to keep an eye on: alternative hosts for protein production in structural biology.

机构信息

Center for Biological Research, Spanish National Research Council (CIB-CSIC), Ramiro de Maeztu 9, 28040 Madrid, Spain.

出版信息

Curr Opin Struct Biol. 2013 Jun;23(3):365-73. doi: 10.1016/j.sbi.2013.02.002. Epub 2013 Mar 5.

Abstract

The conduct of many trials for the successful production of large quantities of pure proteins for structural biology and biotechnology applications, particularly of active, authentically processed enzymes, large eukaryotic multi-subunit complexes and membrane proteins, has spurred the development of recombinant expression systems. Beyond the well-established Escherichia coli, mammalian cell culture and baculovirus-infected insect cell expression systems, a plethora of alternative expression systems has been discovered, engineered, matured and deployed, resulting in crystal, nuclear magnetic resonance, and electron microscopy structures. In this review, we visit alternative expression hosts for structural biology ranging from bacteria and archaea to filamentous and unicellular yeasts and protozoa, with particular emphasis on their applicability to the structural determination of high-value, challenging proteins and complexes.

摘要

为了成功生产大量用于结构生物学和生物技术应用的纯蛋白质,特别是活性的、真实加工的酶、大型真核多亚基复合物和膜蛋白,进行了许多试验,这推动了重组表达系统的发展。除了成熟的大肠杆菌、哺乳动物细胞培养和杆状病毒感染的昆虫细胞表达系统外,还发现、设计、成熟和部署了大量替代表达系统,从而获得了晶体、核磁共振和电子显微镜结构。在这篇综述中,我们访问了结构生物学的替代表达宿主,从细菌和古菌到丝状和单细胞酵母和原生动物,特别强调了它们在确定高价值、具有挑战性的蛋白质和复合物的结构方面的适用性。

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