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

TitleAn allylic/acyclic adenosine nucleoside triphosphate for termination of DNA synthesis by DNA template-dependent polymerases.
Publication TypeJournal Article
Year of Publication1999
AuthorsMartinez, CI, Thoresen, LH, Gibbs, RA, Burgess, K
JournalNucleic Acids Res
Volume27
Issue5
Pagination1271-4
Date Published1999 Mar 01
ISSN0305-1048
KeywordsAdenosine Triphosphate, Base Sequence, Boron Compounds, DNA, DNA Primers, DNA Replication, DNA-Directed DNA Polymerase, Fluorescent Dyes, Kinetics, Substrate Specificity, Templates, Genetic
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.

DOI10.1093/nar/27.5.1271
Alternate JournalNucleic Acids Res.
PubMed ID9973613
PubMed Central IDPMC148311
Grant ListHG01745 / HG / NHGRI NIH HHS / United States