Simple and Easy Synthesis of γ-Amido-dNTPs in Water and Their Polymerase Reaction Properties

Chem Pharm Bull (Tokyo). 2021;69(11):1061-1066. doi: 10.1248/cpb.c21-00497.

Abstract

γ-Amido-modified 2'-deoxynucleoside triphosphates (dNTPs) and nucleoside triphosphates (NTPs) are becoming increasingly important as biological tools. We herein describe the simple and easy synthesis of γ-amido-dNTPs and -NTPs from commercially available corresponding dNTPs and NTPs in a one-pot reaction using water-soluble carbodiimide and ammonia solution. We examined the effects of synthesized γ-amido-dNTPs on the DNA polymerase reaction. The results obtained showed the incorporation of these derivatives into the DNA primer while maintaining nucleobase selectivity; however, their incorporation efficiency by DNA polymerase was lower than that of dNTP. This is the first study to demonstrate the successful synthesis of four sets of γ-amido-dNTPs and clarify their properties.

Keywords: one-pot reaction; polymerase reaction; γ-amido-2′-deoxynucleoside triphosphate.

MeSH terms

  • Ammonia / chemistry
  • Carbodiimides / chemistry
  • Chromatography, High Pressure Liquid
  • DNA-Directed DNA Polymerase / metabolism*
  • Kinetics
  • Nucleotides / chemical synthesis*
  • Polyphosphates / chemistry*
  • Solubility
  • Water

Substances

  • Carbodiimides
  • Nucleotides
  • Polyphosphates
  • Water
  • Ammonia
  • DNA-Directed DNA Polymerase
  • triphosphoric acid