Site-specific discrimination of Cytosine and 5-methylcytosine in duplex DNA by Peptide nucleic acids

J Am Chem Soc. 2002 Sep 4;124(35):10262-3. doi: 10.1021/ja0264955.

Abstract

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.

MeSH terms

  • 5-Methylcytosine
  • Cytosine / analogs & derivatives*
  • Cytosine / analysis
  • Cytosine / chemistry*
  • DNA / analysis
  • DNA / chemistry*
  • Energy Transfer
  • Fluorescein / chemistry
  • Fluorescence
  • Genes, p53
  • Humans
  • Oligonucleotide Probes / chemistry
  • Peptide Nucleic Acids / chemistry*
  • Substrate Specificity

Substances

  • Oligonucleotide Probes
  • Peptide Nucleic Acids
  • 5-Methylcytosine
  • Cytosine
  • DNA
  • Fluorescein