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An allylic/acyclic adenosine nucleoside triphosphate for termination of DNA synthesis by DNA template-dependent
C I Martinez1, L H Thoresen, R A Gibbs
1Department of Chemistry, PO Box 300012, Texas A & M University, College Station, TX 77842-3012, USA.
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.