Nozawa Akira, Nemoto Keiichirou, Nomura Shunsuke, Yamanaka Satoshi, Kido Kohki, Sawasaki Tatsuya
Proteo-Science Center, Ehime University, 3 Bunkyo-cho, Matsuyama, Ehime, 790-8577, Japan.
Methods Mol Biol. 2017;1621:37-46. doi: 10.1007/978-1-4939-7063-6_4.
The wheat germ cell-free protein synthesis system has been used as a eukaryotic protein production system since it was first reported in 1964. Although initially the productivity of this system was not very high, it has now become one of the most versatile protein production systems, thanks to the enhancements made by several groups. In this chapter, we report a protein production method for plant receptor kinases using the wheat cell-free system. We describe a method for the preparation of a cell-free extract from wheat germ, the split-primer PCR method for preparation of transcription templates, and the bilayer cell-free protein synthesis method.
自1964年首次报道以来，小麦胚无细胞蛋白质合成系统一直被用作真核蛋白质生产系统。尽管最初该系统的生产率不是很高，但由于几个研究小组的改进，它现在已成为最通用的蛋白质生产系统之一。在本章中，我们报道了一种使用小麦无细胞系统生产植物受体激酶的蛋白质生产方法。我们描述了从小麦胚制备无细胞提取物的方法、用于制备转录模板的裂分引物PCR方法以及双层无细胞蛋白质合成方法。
