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

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

doi:10.1093/nass/48.1.151

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

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

Authors

Yoshihito Kitamura¹, Junya Yoshida, Wen Chen, Jing-Min Zhou, Jun Sumaoka, Makoto Komiyama

Affiliation

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

PMID: 17150523
DOI: 10.1093/nass/48.1.151

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cerium / metabolism*
DNA / metabolism*
DNA Ligases / metabolism
DNA Restriction Enzymes / metabolism
Edetic Acid / metabolism*
Electrophoresis, Polyacrylamide Gel
Hydrolysis
Oligonucleotides / chemistry*
Phosphates / chemistry*

Substances

Oligonucleotides
Phosphates
Cerium
DNA
Edetic Acid
DNA Restriction Enzymes
DNA Ligases