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γ-酰胺基脱氧核苷三磷酸在水中的简易合成及其聚合酶反应特性

Simple and Easy Synthesis of γ-Amido-dNTPs in Water and Their Polymerase Reaction Properties.

作者信息

Sakurada Takato, Miyahara Ryo, Kawazoe Ryoji, Nagata Yusuke, Kikukawa Yoshiya, Sasaki Shigeki, Taniguchi Yosuke

机构信息

Graduate School of Pharmaceutical Sciences, Kyushu University.

Graduate School of Pharmaceutical Sciences, Nagasaki International University.

出版信息

Chem Pharm Bull (Tokyo). 2021;69(11):1061-1066. doi: 10.1248/cpb.c21-00497.

Abstract

γ-Amido-modified 2'-deoxynucleoside triphosphates (dNTPs) and nucleoside triphosphates (NTPs) are becoming increasingly important as biological tools. We herein describe the simple and easy synthesis of γ-amido-dNTPs and -NTPs from commercially available corresponding dNTPs and NTPs in a one-pot reaction using water-soluble carbodiimide and ammonia solution. We examined the effects of synthesized γ-amido-dNTPs on the DNA polymerase reaction. The results obtained showed the incorporation of these derivatives into the DNA primer while maintaining nucleobase selectivity; however, their incorporation efficiency by DNA polymerase was lower than that of dNTP. This is the first study to demonstrate the successful synthesis of four sets of γ-amido-dNTPs and clarify their properties.

摘要

γ-酰胺基修饰的2'-脱氧核苷三磷酸(dNTPs)和核苷三磷酸(NTPs)作为生物学工具正变得越来越重要。我们在此描述了一种简单易行的合成方法,即使用水溶性碳二亚胺和氨溶液,通过一锅法反应从市售的相应dNTPs和NTPs合成γ-酰胺基-dNTPs和-NTPs。我们研究了合成的γ-酰胺基-dNTPs对DNA聚合酶反应的影响。所得结果表明,这些衍生物在保持碱基选择性的同时被掺入DNA引物中;然而,它们被DNA聚合酶的掺入效率低于dNTP。这是首次证明成功合成四组γ-酰胺基-dNTPs并阐明其性质的研究。

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