Manipulation of DNA through Ce(IV)/EDTA-induced site-selective hydrolysis using phosphate-bearing oligonucleotides.
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.
Kitamura
Yoshihito
Y
Research Center for Advanced Science and Technology, The University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo 153-8904, Japan.
Yoshida
Junya
J
Chen
Wen
W
Zhou
Jing-Min
JM
Sumaoka
Jun
J
Komiyama
Makoto
M
D002563:Cerium / Q000378:metabolism*; D004247:DNA / Q000378:metabolism*; D011088:DNA Ligases / Q000378:metabolism; D004262:DNA Restriction Enzymes / Q000378:metabolism; D004492:Edetic Acid / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D006868:Hydrolysis; D009841:Oligonucleotides / Q000737:chemistry*; D010710:Phosphates / Q000737:chemistry*
D009841:Oligonucleotides; D010710:Phosphates; D002563:Cerium; D004247:DNA; D004492:Edetic Acid; D004262:DNA Restriction Enzymes; D011088:DNA Ligases