无细胞蛋白质合成
翻译系统
翻译(生物学)
蛋白质生物合成
重组DNA
可控性
大肠杆菌
化学
合成生物学
无细胞系统
计算生物学
生物化学
组合化学
生物
信使核糖核酸
酶
数学
体外
基因
应用数学
作者
Yoshihiro Shimizu,Takashi Kanamori,Takuya Ueda
出处
期刊:Methods
[Elsevier]
日期:2005-07-01
卷期号:36 (3): 299-304
被引量:364
标识
DOI:10.1016/j.ymeth.2005.04.006
摘要
We have developed a partially recombinant, cell-free, protein-synthesis system reconstituted solely from those essential elements of the Escherichia coli translation system, termed protein synthesis using recombinant elements (PURE). It provides higher reaction controllability in comparison to crude cell-free protein-synthesis systems for translation studies and biotechnology applications. The PURE system stands out among translation methods in that it provides not only a simple and unique “reverse” purification method of separating the synthesized protein from reaction mixture, but also that the system can be tailor-made according to individual protein requirements. In this paper, two new approaches to obtaining active proteins are described: the use of molecular chaperones, and modification of the reaction conditions. Several possible applications of the PURE system are also discussed.
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