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[RNA连接酶反应的中间产物]

[Intermediate products of the RNA-ligase reaction].

作者信息

Iuodka B A, Snechkute M A, Ianushonite L M, Markutskas A Ia

出版信息

Biokhimiia. 1979 Apr;44(4):599-604.

PMID:35245
Abstract

The first and the third steps of the RNA-ligase reaction were studied. It was shown that the first step of the reaction consists in a formation of an enzyme-adenylate complex. The optimal conditions for this formation were established. Effects of acids, alkali, hydroxylamine and snake venom phosphodiesterase on the complex suggest that the linkage between the protein and adenylic acid may be of a phosphoester or phosphoamide type. Using synthetic adenylic acid pyrophosphates and mononucleotides (oligonucleotides) the RNA-ligase reaction was shown to involve intermediate pyrophosphates. It was found that the simplest pyrophosphates capable to bind to oligonucleotides in the absence of ATP are adenylic acid pyrophosphates, both of ribo- and deoxyribomononucleotides. The RNA-ligase reaction may be used for elongation of oligonucleotides by one definite mononucleotide or for incorporation of the label into the 3'-end of the polynucleotide chain.

摘要

对RNA连接酶反应的第一步和第三步进行了研究。结果表明,反应的第一步在于形成酶 - 腺苷酸复合物。确定了该复合物形成的最佳条件。酸、碱、羟胺和蛇毒磷酸二酯酶对该复合物的影响表明,蛋白质与腺苷酸之间的连接可能是磷酸酯或磷酰胺类型。使用合成的腺苷酸焦磷酸和单核苷酸(寡核苷酸)表明,RNA连接酶反应涉及中间焦磷酸。发现能够在没有ATP的情况下与寡核苷酸结合的最简单焦磷酸是腺苷酸焦磷酸,包括核糖和脱氧核糖单核苷酸的腺苷酸焦磷酸。RNA连接酶反应可用于通过一种特定的单核苷酸延长寡核苷酸,或用于将标记掺入多核苷酸链的3'-末端。

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