Kigawa Takanori
RIKEN Systems and Structural Biology Center, Yokohama, Japan.
Methods Mol Biol. 2010;607:1-10. doi: 10.1007/978-1-60327-331-2_1.
We have been developing and using an Escherichia coli cell extract-based coupled transcription-translation cell-free system. The development includes many different issues such as cell extract preparation, template construction, reaction condition, reaction format, and automation. These developments improved the efficiency, productivity, and throughput of our prokaryotic cell-free system, enabling us to use the system as one of the standard expression methods in our group. Our system certainly has the largest successful applications especially to the protein production for the structure determination, among the existing cell-free protein synthesis systems.
我们一直在开发并使用一种基于大肠杆菌细胞提取物的转录-翻译无细胞偶联系统。该开发过程涉及许多不同的问题，如细胞提取物的制备、模板构建、反应条件、反应形式以及自动化。这些进展提高了我们原核无细胞系统的效率、产量和通量，使我们能够将该系统用作我们团队的标准表达方法之一。在现有的无细胞蛋白质合成系统中，我们的系统在用于结构测定的蛋白质生产方面无疑拥有最为成功的应用。
