含2'-脱氧-6-硫代肌苷的寡脱氧核苷酸的简便合成

Convenient synthesis of oligodeoxynucleotides containing 2'-deoxy-6-thioinosine.

作者信息

Beuck Christine, Weinhold Elmar

机构信息

Institut für Organische Chemie der RWTH Aachen, Aachen, Germany.

出版信息

Nucleosides Nucleotides Nucleic Acids. 2003 May-Aug;22(5-8):635-9. doi: 10.1081/NCN-120021969.

DOI:10.1081/NCN-120021969

PMID:14565242

Abstract

A facile synthesis of oligodeoxynucleotides (ODN) containing 2'-deoxy-6-thioinosine (dI6S) based on the convertible nucleoside O6-phenyl-2'-deoxyinosine is presented. After standard solid-phase DNA synthesis and removal of the cyanoethyl protecting groups with DBU treatment with aqueous sodium hydrogen sulfide introduces the sulfur functionality, deprotects the other nucleobases and cleaves the ODN from the solid support in a one-pot reaction. In addition, the extinction coefficient of 2'-deoxy-6-thioinosine is determined by enzymatic fragmentation of the resulting ODN in the presence of adenosine deaminase.

摘要

本文介绍了一种基于可转化核苷O6-苯基-2'-脱氧肌苷的含2'-脱氧-6-硫代肌苷（dI6S）的寡脱氧核苷酸（ODN）的简便合成方法。经过标准的固相DNA合成，并用DBU处理除去氰乙基保护基后，用硫化氢水溶液引入硫官能团，使其他核苷酸碱基脱保护，并在一锅反应中将ODN从固相载体上切割下来。此外，通过在腺苷脱氨酶存在下对所得ODN进行酶促片段化来测定2'-脱氧-6-硫代肌苷的消光系数。