利用RNA连接酶，以活化的单核苷酸及其类似物在3'方向上延伸寡核苷酸。
Elongation of oligonucleotides in the 3'-direction with activated mononucleotides and their analogs using RNA ligase.
作者信息
Ohtsuka E, Miyake T, Nagao K, Uemura H, Nishikawa S, Sugiura M, Ikehara M
出版信息
Nucleic Acids Res. 1980 Feb 11;8(3):601-10. doi: 10.1093/nar/8.3.601.
Abstract
P1-Adenosine 5'-P2-2',3'-ethoxymethylidene nucleosides [A(5')ppN(Em)] from four common nucleosides have been prepared and used for single addition of nucleotides to elongate oligonucleotide chains in the 3'-direction in RNA ligase reaction. U-U-C, T-U-C and A-C-C were used as acceptors. Structural dependence in these acceptors was found to be smaller compared to joining reactions between oligonucleotides. Adenosine analogs including 8-bromo-, 2'-fluoro-, 2'-azido-, 8,2'-O-cyclo-, 8,2'-S-cyclo-adenosine, arabinosyladenine and 2'-deoxyadenosine were added to the 3'-end of A-C-C by adenylation chemically followed by joining with RNA ligase. Symmetrical 5'-pyrophosphates of 8-bromo-, 2'-fluoro- and 2'-azido-adenosine were not recognized as donor substrates.
摘要
已制备了来自四种常见核苷的P1-腺苷5'-P2-2',3'-乙氧基亚甲基核苷[A(5')ppN(Em)]，并将其用于在RNA连接酶反应中以3'方向单加核苷酸来延长寡核苷酸链。U-U-C、T-U-C和A-C-C用作受体。与寡核苷酸之间的连接反应相比，发现这些受体中的结构依赖性较小。通过化学腺苷化将包括8-溴-、2'-氟-、2'-叠氮基-、8,2'-O-环-、8,2'-S-环-腺苷、阿拉伯糖基腺嘌呤和2'-脱氧腺苷在内的腺苷类似物添加到A-C-C的3'末端，然后与RNA连接酶连接。8-溴-、2'-氟-和2'-叠氮基-腺苷的对称5'-焦磷酸酯未被识别为供体底物。
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本文引用的文献
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