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Nucleic Acids Symposium Series 2004 48(1):151-152; doi:10.1093/nass/48.1.151
© 2004 by Oxford University Press
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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 and Makoto Komiyama

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

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.


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