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METHOD FOR EFFICIENT SYNTHESIS OF PROTEIN HAVING LABELED N-TERMINAL AMINO ACID RESIDUE

机译：带有标签的N-末端氨基酸残基的蛋白质的高效合成方法

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摘要

An N-terminus-labeling method is a method for synthesis of a protein having a specifically labeled α-amino group in an N-terminal methionine residue. In a conventional N-terminus-labeling method, the degree of efficiency of labeling (i.e., the percentage of labeled proteins relative to the total synthesized proteins) has been as extremely small as 1-2%. The degree of efficiency of labeling can be significantly improved by (1) using a tRNA-mixed solution which comprises substantially only stretched tRNA having initiation tRNA removed therefrom and utilizing a synthesis system (an in vitro transcription/translation system) which does not contain methyonyl tRNA transformylase and 10-formyl-5,6,7,8-tetrahydrofolic acid; (2) isolating/purifying labeled methyonyl initiation tRNA; and/or (3) controlling the time required for the labeled-protein synthesis reaction.

机译：N末端标记法是合成在N末端甲硫氨酸残基中具有被特定标记的α-氨基的蛋白质的方法。在常规的N末端标记方法中，标记的效率（即，标记的蛋白质相对于总的合成蛋白质的百分比）非常小，为1-2％。可以通过以下方法显着提高标记效率：（1）使用基本上只包含拉伸的tRNA的tRNA混合溶液，其中去除了起始tRNA，并使用不包含甲炔基的合成系统（体外转录/翻译系统） tRNA转化酶和10-甲酰基-5,6,7,8-四氢叶酸; （2）分离/纯化标记的甲氧甲基起始tRNA;和/或（3）控制标记蛋白质合成反应所需的时间。

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公开/公告号WO2007058376A1

专利类型
公开/公告日2007-05-24

原文格式PDF
申请/专利权人 POST GENOME INSTITUTE CO. LTD.;GIFU UNIVERSITY;KANAMORI TAKASHI;NISHIKAWA KAZUYA;YOKOGAWA TAKASHI;OHNO SATOSHI;
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申请/专利号WO2006JP323378
发明设计人 NISHIKAWA KAZUYA;YOKOGAWA TAKASHI;OHNO SATOSHI;KANAMORI TAKASHI;
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申请日2006-11-16
分类号C12P21/00;C12N15/09;
国家 WO
入库时间 2022-08-21 20:49:40

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