Manipulation of DNA through Ce(IV)/EDTA-induced site-selective hydrolysis using phosphate-bearing oligonucleotides.

作者信息

Kitamura Yoshihito, Yoshida Junya, Chen Wen, Zhou Jing-Min, Sumaoka Jun, Komiyama Makoto

机构信息

Research Center for Advanced Science and Technology, The University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo 153-8904, Japan.

出版信息

Nucleic Acids Symp Ser (Oxf). 2004(48):151-2. doi: 10.1093/nass/48.1.151.

DOI:10.1093/nass/48.1.151

PMID:17150523

Abstract

By using two oligonucleotides bearing a monophosphate group at the terminus and placing them at both edges of gap-site formed in substrate DNA, this site was preferentially and efficiently hydrolyzed by Ce(IV)/EDTA. In this study, site-selective hydrolysis of gap-site in long substrate DNA (274-mer) was investigated. When two monophosphate groups were placed to both edges of 5-base gap, predetermined site was selectively hydrolyzed by Ce(IV)/EDTA. Furthermore, desired recombinant DNA was prepared from this fragment and foreign DNA using T4 DNA ligase.

摘要