Electron microscopy visualization of oligonucleotide binding to duplex DNA via triplex formation

J Mol Biol. 1993 Mar 20;230(2):379-83. doi: 10.1006/jmbi.1993.1154.

Abstract

Using biotinylated oligonucleotides and streptavidin as a marker, we have visualized, with the help of electron microscopy, the triplex formation. We used the natural homopurine-homopyrimidine sequence from human papillomavirus 16 cloned within a plasmid. Under conditions favouring the formation of pyrimidine-purine-pyrimidine triplex the corresponding pyrimidine oligonucleotide formed a complex with the insert and this complex was detected by electron microscopy. Similarly, under conditions favouring the formation of pyrimidine-purine-purine triplex the corresponding purine oligonucleotide formed a stable complex detected by electron microscopy. In both cases the complexes we observed exhibited remarkable sequence specificity. Near 80% of DNA molecules carried the streptavidin marker in the correct position and very few cases of non-specific binding were detected. We conclude that the triplex mode of recognition may provide very efficient sequence-specific markers for electron microscopy of DNA.

Publication types

  • Comparative Study

MeSH terms

  • Base Sequence
  • Binding Sites
  • DNA / chemistry*
  • DNA / ultrastructure
  • Microscopy, Electron / methods
  • Models, Structural
  • Molecular Sequence Data
  • Nucleic Acid Conformation
  • Oligodeoxyribonucleotides / chemistry*
  • Structure-Activity Relationship

Substances

  • Oligodeoxyribonucleotides
  • DNA