Shimizu Yoshihiro, Ueda Takuya
The Department of Medical Genome Sciences, Graduate School of Frontier Sciences, The University of Tokyo, Kashiwa, Chiba, Japan.
Methods Mol Biol. 2010;607:11-21. doi: 10.1007/978-1-60327-331-2_2.
The Escherichia coli-based reconstituted cell-free protein synthesis system, which we named the PURE (Protein synthesis Using Recombinant Elements) system, provides several advantages compared with the conventional cell-extract-based system. Stability of RNA or protein is highly improved because of the lack of harmful degradation enzymes. The system can be easily engineered according to purposes or the proteins to be synthesized, by manipulating the components in the system. In this chapter, we describe the construction and exploitation of the PURE system. Methods for preparing and assembling the components composing the PURE system for the protein synthesis reaction are shown.
基于大肠杆菌的无细胞重组蛋白质合成系统,我们将其命名为PURE(使用重组元件的蛋白质合成)系统,与传统的基于细胞提取物的系统相比具有若干优势。由于缺乏有害的降解酶,RNA或蛋白质的稳定性得到了极大提高。通过操纵系统中的组分,该系统可根据目的或待合成的蛋白质轻松进行改造。在本章中,我们描述了PURE系统的构建与应用。展示了制备和组装用于蛋白质合成反应的PURE系统各组分的方法。