Synthesis and properties of oligonucleotides containing 5-formyl-2'-deoxycytidine: in vitro DNA polymerase reactions on DNA templates containing 5-formyl-2'-deoxycytidine

Nucleic Acids Res. 2001 Jun 15;29(12):2456-63. doi: 10.1093/nar/29.12.2456.


Oligodeoxynucleotides (ODNs) containing 5-formyl-2'-deoxycytidine (fC) were synthesized by the phosphoramidite method and subsequent oxidation with sodium periodate. The stabilities of duplexes containing A, G, C or T opposite fC were studied by thermal denaturation. It was found that fC:A, fC:C or fC:T base pairs significantly reduce the thermal stabilities of duplexes. Next, single nucleotide insertion reactions were performed using ODNs containing fC as templates and the Klenow fragment of Escherichia coli DNA polymerase I. It was found that: (i) insertion of dGMP opposite fC appears to be less efficient relative to insertion opposite 5-methyl-2'-deoxycytidine (mC); (ii) dAMP is misincorporated more frequently opposite fC than mC, although the frequency of misincorporation seems to be dependent on the sequence; (iii) TMP is misincorporated more frequently opposite fC than mC. These results suggest that fC may induce the transition mutation C.G-->T.A and the transversion mutation C.G-->A.T during DNA synthesis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Pairing
  • Base Sequence
  • DNA / biosynthesis*
  • DNA / chemistry
  • DNA / genetics
  • DNA Polymerase I / metabolism*
  • DNA Primers / genetics
  • DNA Replication*
  • Deoxycytidine / analogs & derivatives
  • Deoxycytidine / chemistry
  • Deoxycytidine / metabolism*
  • Escherichia coli / enzymology
  • Kinetics
  • Magnetic Resonance Spectroscopy
  • Mutagenesis / genetics*
  • Nucleic Acid Denaturation / radiation effects
  • Nucleic Acid Hybridization
  • Oligonucleotides / chemical synthesis*
  • Oligonucleotides / chemistry
  • Oligonucleotides / genetics
  • Oligonucleotides / metabolism*
  • Periodic Acid / metabolism
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
  • Temperature
  • Templates, Genetic
  • Thermodynamics
  • Ultraviolet Rays


  • 5-formyl-2'-deoxycytidine
  • DNA Primers
  • Oligonucleotides
  • Deoxycytidine
  • Periodic Acid
  • DNA
  • metaperiodate
  • DNA Polymerase I