Protein synthesis by pure translation systems
Protein synthesis by pure translation systems
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DOI：
10.1016/j.ymeth.2005.04.006
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发表时间：
2005-07-01
期刊：
影响因子：
4.8
通讯作者：
Ueda, T
中科院分区：
文献类型：
--
作者：
Shimizu, Y;Kanamori, T;Ueda, T
我们已经开发了一种部分重组的，无细胞的，蛋白质合成系统，仅从大肠杆菌翻译系统的基本元素，称为蛋白质合成使用重组元件（纯）重建。与用于翻译研究和生物技术应用的粗无细胞蛋白质合成系统相比，它提供了更高的反应可控性。PURE系统在翻译方法中脱颖而出，因为它不仅提供了从反应混合物中分离合成蛋白质的简单而独特的“反向”纯化方法，而且该系统可以根据单个蛋白质的需求量身定制。本文介绍了两种获得活性蛋白的新方法：分子伴侣的使用和反应条件的改变。PURE系统的几个可能的应用进行了讨论。(c)2005年爱思唯尔公司All rights reserved.
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. (c) 2005 Elsevier Inc. All rights reserved.