Oligonucleotide bearing ethylenediamine-N,N,N′-triacetates for gap-selective DNA hydrolysis by Ce4+/EDTA
Oligonucleotide bearing ethylenediamine-N,N,N′-triacetates for gap-selective DNA hydrolysis by Ce4+/EDTA
复制标题
DOI：
10.1002/cbic.200400220
复制
发表时间：
2005-01-01
期刊：
影响因子：
3.2
通讯作者：
Yamamoto, Y
中科院分区：
文献类型：
--
作者：
Komiyama, M;Arishima, H;Yamamoto, Y
利用含有乙二胺-N，N，N'-三乙酸基团的两种寡核苷酸作为添加剂，在底物DNA的预定位置形成间隙位点。用Ce4+/EDTA络合物在pH 7.0和37℃下处理这些体系；间隙处的磷酸二酯键被选择性水解。乙二胺三乙酸基的引入极大地促进了DNA的断裂，并且随着基数的增加，断裂效率也随之提高。当三个乙二胺三乙酸基团连续放置在间隙的两边时，即使是一个碱基的间隙也能成功水解，尽管在没有这些基团的情况下裂解很小。位点选择性断裂也可以在更高的温度下实现，而不会显著损失位点选择性。
With the use of two oligonucleotides bearing ethylenediamine-N,N,N'-triacetate groups as additives, gap sites were formed at predetermined sites in substrate DNA. Upon treating these systems with a Ce4+/EDTA complex at pH 7.0 and 37 degreesC; the phosphodiester linkages at the gap site were selectively hydrolyzed. The DNA scission was greatly promoted by the introduction of ethylenediaminetriacetate groups, and the scission efficiency increased as the number of these groups increased. Even a one-base gap was successfully hydrolyzed when three ethylene-diaminetriacetate groups were placed consecutively at both edges of the gap, although the scission was minimal in the absence of these groups. The site-selective scission could be also achieved at higher temperatures without any significant loss of site-selectivity.