Application of cerium(IV)/EDTA complex for future biotechnology
Application of cerium(IV)/EDTA complex for future biotechnology
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DOI：
10.1016/j.jallcom.2004.12.075
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发表时间：
2006-02
影响因子：
6.2
通讯作者：
J. Sumaoka;Wen Chen;Yoshihito Kitamura;T. Tomita;J. Yoshida;M. Komiyama
中科院分区：
文献类型：
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作者：
J. Sumaoka;Wen Chen;Yoshihito Kitamura;T. Tomita;J. Yoshida;M. Komiyama
制备了一种新的单链DNA定点水解人工体系。通过使用在末端带有单磷酸基团的两种寡核苷酸添加剂，在底物DNA中的预定位置形成缺口结构。在pH 7.0和37°C下，Ce（IV）/EDTA络合物（EDTA，乙二胺-N，N，N′，N′-四乙酸盐）可有效地选择性水解差距中的磷酸二酯键。此外，通过位点选择性切断形成的片段通过使用T4 DNA连接酶与各种寡核苷酸连接，产生所需的重组DNA。成功地获得了一种在生物技术中操纵单链DNA的新工具。
A new artificial system for site-selective hydrolysis of single-stranded DNAs was prepared. By using two oligonucleotide additives that bear a monophosphate group at the termini, gap structures were formed at predetermined positions in substrate DNA. The phosphodiester linkages in the gap were efficiently and selectively hydrolyzed by Ce(IV)/EDTA complex (EDTA, ethylenediamine-N,N,N′,N′-tetraacetate) at pH 7.0 and 37°C. Furthermore, the fragments formed by the site-selective scission were connected with various oligonucleotides by using T4 DNA ligase, producing desired recombinant DNAs. A new tool for manipulation of single-stranded DNA in biotechnology has been successfully obtained.