An allylic/acyclic adenosine nucleoside triphosphate for termination of DNA synthesis by DNA template-dependent polymerases.
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.
Martinez
C I
CI
Department of Chemistry, PO Box 300012, Texas A & M University, College Station, TX 77842-3012, USA.
Thoresen
L H
LH
Gibbs
R A
RA
Burgess
K
K
D000255:Adenosine Triphosphate / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000378:metabolism / Q000494:pharmacology; D001483:Base Sequence; D001896:Boron Compounds; D004247:DNA / Q000378:metabolism; D017931:DNA Primers; D004261:DNA Replication / Q000187:drug effects*; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism*; D005456:Fluorescent Dyes; D007700:Kinetics; D013379:Substrate Specificity; D013698:Templates, Genetic