Department of Chemistry and Molecular Biology, University of Gothenburg, Gothenburg, Sweden.
Swedish NMR Centre, University of Gothenburg, Gothenburg, Sweden.
Methods Mol Biol. 2020;2141:233-245. doi: 10.1007/978-1-0716-0524-0_11.
Cell-free protein synthesis (CFPS) is an established method to produce recombinant proteins and has been used in a wide variety of applications. The use of CFPS has almost from the onset been favorably linked to the production of isotopically labelled proteins for NMR spectroscopy as the resulting labelling of the produced protein is defined by the chosen amino acids during reaction setup. Here we describe how to set up production and isotopic labelling of small intrinsically disordered proteins (IDPs) for NMR spectroscopy applications using an E. coli-based CFPS system in batch mode.
无细胞蛋白质合成（CFPS）是一种生产重组蛋白的成熟方法，已广泛应用于各种应用中。CFPS 的使用几乎从一开始就与用于 NMR 光谱学的同位素标记蛋白质的生产有利地联系在一起，因为所产生的蛋白质标记是由反应设置过程中选择的氨基酸定义的。在这里，我们描述了如何使用基于大肠杆菌的 CFPS 系统在批量模式下为 NMR 光谱学应用设置生产和同位素标记小分子无序蛋白质（IDP）。
