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C-terminal modified protein and method for producing the same, modifying agent and translation template used for producing C-terminal modified protein, and method for detecting protein interaction with use of C-terminal modified protein

机译:C末端修饰蛋白及其制造方法,用于产生C末端修饰蛋白的修饰剂和翻译模板,以及使用该C末端修饰蛋白检测蛋白质相互作用的方法

摘要

A translation template comprising an ORF region coding for a protein, a 5′ untranslated region comprising a transcription promoter and a translation enhancer and locating on the 5′ side of the ORF region, and a 3′ end region comprising a poly-A sequence and locating on the 3′ side of the ORF region, is expressed in a translation system in the presence of an agent for modifying a C-terminal of a protein, which comprises an acceptor portion having a group capable of binding to a protein through a transpeptidation reaction in a protein translation system and a modifying portion comprising a nonradioactive modifying substance linked to the acceptor portion via a nucleotide linker, to cause protein synthesis and the synthesized protein is purified. Thus, the yield of modified protein in a method of modifying C-terminal of protein is improved and detection of protein interaction based on various intermolecular interaction detection methods is realized at an improved level.
机译:一种翻译模板,其包含编码蛋白质的ORF区,包含转录启动子和翻译增强子并位于ORF区5'侧的5'非翻译区以及包含poly-A序列和位于ORF区3'侧的蛋白在修饰蛋白质C端的试剂存在下在翻译系统中表达,该试剂包含一个受体部分,该受体部分具有能够通过转肽作用与蛋白质结合的基团在蛋白质翻译系统中进行的反应和包含非放射性修饰物质的修饰部分通过核苷酸接头与受体部分连接,从而引起蛋白质合成,并纯化合成的蛋白质。因此,提高了在修饰蛋白质的C末端的方法中修饰的蛋白质的产率,并且以改进的水平实现了基于各种分子间相互作用检测方法的蛋白质相互作用的检测。

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