Site-specific discrimination of Cytosine and 5-methylcytosine in duplex DNA by Peptide nucleic acids.
For site-specific discrimination of cytosine (C) and 5-methylcytosine (mC) in duplex DNA, we developed a new method using peptide nucleic acids (PNAs). The combination of a PNA-assisted DNA displacement complex and a fluorescein-labeled probe oligomer allowed the detection of mC at the defined sites in target DNA using a restriction enzyme. After treatment of the complex with a restriction enzyme, strong fluorescence emission was observed for the complex containing C at the target site, whereas the fluorescence intensity for the complex containing mC was extremely weak.
Okamoto
Akimitsu
A
Department of Synthetic Chemistry and Biological Chemistry, Faculty of Engineering, Kyoto University, and SORST, Japan Science and Technology Corporation, Kyoto 606-8501, Japan.
Tanabe
Kazuhito
K
Saito
Isao
I
D044503:5-Methylcytosine; D003596:Cytosine / Q000031:analogs & derivatives* / Q000032:analysis / Q000737:chemistry*; D004247:DNA / Q000032:analysis / Q000737:chemistry*; D004735:Energy Transfer; D019793:Fluorescein / Q000737:chemistry; D005453:Fluorescence; D016158:Genes, p53; D006801:Humans; D015345:Oligonucleotide Probes / Q000737:chemistry; D020135:Peptide Nucleic Acids / Q000737:chemistry*; D013379:Substrate Specificity
D015345:Oligonucleotide Probes; D020135:Peptide Nucleic Acids; D044503:5-Methylcytosine; D003596:Cytosine; D004247:DNA; D019793:Fluorescein