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

Yoshiaki Kitamura | Zendy

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

Author(s) -

Yoshiaki Kitamura

Publication year - 2004

Publication title -

nucleic acids symposium series

Language(s) - English

Resource type - Journals

eISSN - 1746-8272

pISSN - 0261-3166

DOI - 10.1093/nass/48.1.151

Subject(s) - dna , oligonucleotide , hydrolysis , dna ligase , chemistry , substrate (aquarium) , phosphate , recombinant dna , active site , stereochemistry , biochemistry , enzyme , biology , gene , ecology

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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