[Photoreactive dTTP analogues as substrates for thermostable DNA polymerase from Thermus thermophilus B35]

Bioorg Khim. 2004 Jul-Aug;30(4):369-74. doi: 10.1023/b:rubi.0000037258.51646.76.
[Article in Russian]


Substrate properties of several dTTP analogues bearing a photoreactive 2-nitro-5-azidobenzoyl (NAB) group attached at position 5 of uracil through linkers of various lengths, dTTP-NAB-x-dUTP (where x = 2, 4, 7-13 is the number of atoms in the linker), were studied. All the analogues are substrates for thermostable Thermus thermophilus B35 DNA polymerase in the elongation reaction of the 5'-32P-labeled primer-template complex. The kinetic parameters of some of the analogues were determined and compared with those of natural dTTP. It was shown that an increase in the linker length results in a higher efficiency of the analogue. The incorporation of NAB-x-dUMP residues into the 3'-primer end did not impede a further elongation of the chain in the presence of natural dNTP.

Publication types

  • English Abstract

MeSH terms

  • Azides / chemical synthesis
  • Azides / chemistry
  • DNA-Directed DNA Polymerase / chemistry*
  • Deoxyuracil Nucleotides / chemical synthesis
  • Deoxyuracil Nucleotides / chemistry*
  • Kinetics
  • Nitroso Compounds / chemical synthesis
  • Nitroso Compounds / chemistry
  • Photochemistry
  • Structure-Activity Relationship
  • Thermus thermophilus / enzymology*
  • Thymine Nucleotides / chemistry*


  • Azides
  • Deoxyuracil Nucleotides
  • Nitroso Compounds
  • Thymine Nucleotides
  • DNA polymerase, Thermus thermophilus
  • DNA-Directed DNA Polymerase
  • thymidine 5'-triphosphate