Shirokov Vladimir A, Kommer Aigar, Kolb Vyacheslav A, Spirin Alexander S
Institute of Protein Research, Russian Academy of Sciences, Pushchino, Moscow, Russia.
Methods Mol Biol. 2007;375:19-55. doi: 10.1007/978-1-59745-388-2_2.
Protein synthesis in cell-free systems is an emerging technology already competing with in vivo expression methods. In this chapter the basic principles of continuous-exchange protein synthesizing systems, and protocols for Escherichia coli and wheat germ translation and transcription-translation systems are described. The ways to improve substrate supply in cell-free systems and mRNA design for eukaryotic system are discussed. Correct folding of the synthesized protein is demonstrated and discussed in detail.
无细胞系统中的蛋白质合成是一项新兴技术，已经在与体内表达方法竞争。在本章中，将描述连续交换蛋白质合成系统的基本原理，以及大肠杆菌和小麦胚芽翻译及转录-翻译系统的实验方案。还将讨论改善无细胞系统中底物供应的方法以及真核系统的mRNA设计。将详细展示和讨论合成蛋白质的正确折叠。
