An allylic/acyclic adenosine nucleoside triphosphate for termination of DNA synthesis by DNA template-dependent polymerases

Nucleic Acids Res. 1999 Mar 1;27(5):1271-4. doi: 10.1093/nar/27.5.1271.

Abstract

An allylic adenosine triphosphate analog (AATP) was tested as a substrate for commercially available DNA polymerases. All but one of the enzymes assayed incorporated AATP opposite thymidine (T) with concomitant termination of the elongation reaction. A concentration of only 1 microM was sufficient for complete termination of the polymerization reaction for a short template mediated by Ampli Taq DNA polymerase FS (Taq FS). This result suggests that AATP could be used as a 2',3'-dideoxyadenosine-5'-triphosphate (ddA) surrogate. Kinetics of incorporation revealed that AATP was 48 times less efficiently incorporated than ddA. Furthermore, AATP was used in dye-primer sequencing as a substitute for ddA.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adenosine Triphosphate / analogs & derivatives*
  • Adenosine Triphosphate / chemical synthesis
  • Adenosine Triphosphate / metabolism
  • Adenosine Triphosphate / pharmacology
  • Base Sequence
  • Boron Compounds
  • DNA / metabolism
  • DNA Primers
  • DNA Replication / drug effects*
  • DNA-Directed DNA Polymerase / metabolism*
  • Fluorescent Dyes
  • Kinetics
  • Substrate Specificity
  • Templates, Genetic

Substances

  • 4,4-difluoro-4-bora-3a,4a-diaza-s-indacene
  • 4-adeninyl-2-(hydroxymethyl)-2-butenyl triphosphate
  • Boron Compounds
  • DNA Primers
  • Fluorescent Dyes
  • Adenosine Triphosphate
  • DNA
  • DNA-Directed DNA Polymerase