The reaction of cytosine with bisulfite by base flipping from the duplex.
Methylation of cytosine at the C5-position in DNA plays a major role in epigenetic gene control. The detection of 5-methylcytosine was designed with the base flipping from the duplex using NC and CGG/CGG triad. Using bisulfite and hydroxylamine, the reaction of cytosine in 11-mer duplex produced the adduct of bisulfite and hydroxylamine, which was isolated by HPLC and identified by MALDI-TOF MS. The duplex containing (5m)C in (5m)CGG/(5m)CGG did not react with bisulfite and hydroxylamine under the same conditions. Further this method enables cytosine adjacent to mismatch guanine to react with bisulfite and hydroxylamine selectively.
Oka
Yoshimi
Y
Institute of Scientific and Industrial Research, Osaka University, 8-1 Mihogaoka, Ibaraki 567-0047, Japan.
Peng
Tao
T
Takei
Fumie
F
Nakatani
Kazuhiko
K
D044503:5-Methylcytosine / Q000032:analysis*; D020137:Base Pair Mismatch; D002219:Carbamates / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid; D003596:Cytosine / Q000737:chemistry*; D004247:DNA / Q000737:chemistry*; D019811:Hydroxylamine / Q000737:chemistry; D009287:Naphthyridines / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013447:Sulfites / Q000737:chemistry*
D002219:Carbamates; D009287:Naphthyridines; D013447:Sulfites; D019811:Hydroxylamine; D044503:5-Methylcytosine; D003596:Cytosine; D004247:DNA; C042345:hydrogen sulfite