Oligonucleotide bearing ethylenediamine-N,N,N'-Triacetates for gap-selective DNA hydrolysis by Ce4+/EDTA.
With the use of two oligonucleotides bearing ethylenediamine-N,N,N'-triacetate groups as additives, gap sites were formed at predetermined sites in substrate DNA. Upon treating these systems with a Ce(4+)/EDTA complex at pH 7.0 and 37 degrees C, the phosphodiester linkages at the gap site were selectively hydrolyzed. The DNA scission was greatly promoted by the introduction of ethylenediaminetriacetate groups, and the scission efficiency increased as the number of these groups increased. Even a one-base gap was successfully hydrolyzed when three ethylenediaminetriacetate groups were placed consecutively at both edges of the gap, although the scission was minimal in the absence of these groups. The site-selective scission could be also achieved at higher temperatures without any significant loss of site-selectivity.
Komiyama
Makoto
M
Research Center for Advanced Science and Technology, The University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo, 153-8904, Japan. komiyama@mkomi.rcast.u-tokyo.ac.jp
Arishima
Hiroyuki
H
Yokoyama
Masahito
M
Kitamura
Yoshihito
Y
Yamamoto
Yoji
Y
D001483:Base Sequence; D002586:Cesium / Q000737:chemistry* / Q000378:metabolism; D002614:Chelating Agents / Q000737:chemistry*; D004247:DNA / Q000378:metabolism*; D004492:Edetic Acid / Q000737:chemistry* / Q000378:metabolism; D006868:Hydrolysis; D008969:Molecular Sequence Data; D009841:Oligonucleotides / Q000737:chemistry* / Q000378:metabolism
D002614:Chelating Agents; D009841:Oligonucleotides; D002586:Cesium; D004247:DNA; D004492:Edetic Acid