Preparation method for Escherichia coliS30 extracts completely dependent upon tRNA addition to catalyze cell-free protein synthesis
Preparation method for Escherichia coliS30 extracts completely dependent upon tRNA addition to catalyze cell-free protein synthesis
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DOI：
10.1007/bf02932309
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发表时间：
2006-09-01
影响因子：
3.2
通讯作者：
Kim, Dong-Myung
中科院分区：
文献类型：
--
作者：
Ahn, Jin-Ho;Hwang, Mi-Yeon;Kim, Dong-Myung
已经开发出一种去除大肠杆菌 S30 提取物内源 tRNA 的简单方法。用水平衡的乙醇胺-Sepharose (R) 柱选择性捕获大肠杆菌 S30 提取物中的 tRNA 分子。结果，通过该柱过滤的 S30 提取物完全依赖于添加外源 tRNA 来介导无细胞蛋白质合成反应。我们预计所开发和描述的程序对于旨在将非天然氨基酸引入蛋白质分子的体外抑制反应研究特别有用。
A simple method for depleting E. coli S30 extracts of endogenous tRNA has been developed. An ethanolamine-Sepharose (R) column equilibrated with water selectively captured the tRNA molecules in E. coli S30 extracts. As a result, S30 extracts filtered through this column became completely dependent upon the addition of exogenous tRNA to mediate cell-free protein synthesis reactions. We anticipate that the procedures developed and described will be particularly useful for in vitro suppression reaction studies designed to introduce unnatural amino acids into protein molecules.