A bilayer cell-free protein synthesis system for high-throughput screening of gene products
A bilayer cell-free protein synthesis system for high-throughput screening of gene products
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DOI：
10.1016/s0014-5793(02)02329-3
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发表时间：
2002-03-06
期刊：
影响因子：
3.5
通讯作者：
Endo, Y
中科院分区：
文献类型：
--
作者：
Sawasaki, T;Hasegawa, Y;Endo, Y
描述了一种高通量无细胞蛋白质合成方法。该方法是基于一个双层扩散系统，使连续供应的基板，连同小的副产物的连续去除，通过翻译混合物和基板混合物之间的相。通过使用多滴定板，该系统在延长的时间内起作用，因此产生的产量是类似分批模式反应的10倍以上。将此方法与我们开发的麦胚无细胞翻译系统相结合，该系统可以产生足够进行功能分析的大量蛋白质。这种新的基于双层的无细胞蛋白质合成系统以其简单、最短时间和低成本可能是后基因组时代有用的实用方法。(C)2002年欧洲生物化学学会联合会。由Elsevier Science B.V.出版，版权所有。
A high-throughput cell-free protein synthesis method has been described. The methodology is based on a bilayer diffusion system that enables the continuous supply of substrates, together with the continuous removal of small byproducts, through a phase between the translation mixture and substrate mixture. With the use of a multititer plate the system was functional for a prolonged time, and as a consequence yielded more than 10 times that of the similar batch-mode reaction. Combining this method with a wheat germ cell-free translation system developed by us, the system could produce a large amount of protein sufficient for carrying out functional analyses. This novel bilayer-based cell-free protein synthesis system with its simplicity, minimum time and low cost may be useful practical methodology in the post-genome era. (C) 2002 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved.