Takai Kazuyuki, Endo Yaeta
Cell-Free Science and Technology Research Center and Venture Business Laboratory, Ehime University, Ehime, Japan.
Methods Mol Biol. 2010;607:23-30. doi: 10.1007/978-1-60327-331-2_3.
The wheat-germ cell-free protein synthesis system had been one of the most efficient eukaryotic cell-free systems since it was first developed in 1964. However, radio-labeled amino acids had long been essential for detection of the products. Since the discovery of a method for prevention of the contamination by a protein synthesis inhibitor originated from endosperm, the wheat cell-free system has found a wide variety of applications in postgenomic high-throughput screening, structural biology, medicine, and so on. In this chapter, we describe a method for preparation of the cell-free extract and a standard protein synthesis method, as the methods for the applications are found in later chapters.
自1964年首次开发以来，小麦胚无细胞蛋白质合成系统一直是最有效的真核无细胞系统之一。然而，放射性标记氨基酸长期以来一直是检测产物所必需的。自从发现一种防止胚乳来源的蛋白质合成抑制剂污染的方法以来，小麦无细胞系统在基因组后高通量筛选、结构生物学、医学等领域有了广泛的应用。在本章中，我们描述了一种无细胞提取物的制备方法和一种标准蛋白质合成方法，因为应用方法将在后续章节中介绍。
