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.
Department of Synthetic Chemistry and Biological Chemistry, Faculty of Engineering, Kyoto University, and SORST, Japan Science and Technology Corporation, Kyoto 606-8501, Japan.
Akimitsu
A
Okamoto
Kazuhito
K
Tanabe
Isao
I
Saito
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