N7-cyanoborane-2'-deoxyguanosine 5'-triphosphate is a good substrate for DNA polymerase.

Published

Journal Article

The 5'-triphosphate of the boronated nucleoside analog N7-cyanoborane-2'-deoxyguanosine (7bdGTP) was synthesized, and a series of experiments was initiated to assess the potential of the compound to serve as a substrate for DNA polymerases. We show here that 7bdGTP can be incorporated into DNA by Sequenase. The resulting hemiboronated extension products are resistant to cleavage by treatment with either DMS and heat or a number of restriction enzymes. Further, in the polymerase chain reaction, 7bdGTP can be utilized as a substrate for Taq polymerase. Finally, by kinetic analysis, we have found that 7bdGTP is a more efficient substrate for exonuclease-free Klenow than normal dGTP. Thus, the introduction of a cyanoborane moiety to the N7 position of dGTP results in a nucleotide that is accepted in lieu of normal dGTP by a number of DNA polymerases.

Full Text

Duke Authors

Cited Authors

  • Porter, KW; Tomasz, J; Huang, F; Sood, A; Shaw, BR

Published Date

  • September 1995

Published In

Volume / Issue

  • 34 / 37

Start / End Page

  • 11963 - 11969

PubMed ID

  • 7547933

Pubmed Central ID

  • 7547933

Electronic International Standard Serial Number (EISSN)

  • 1520-4995

International Standard Serial Number (ISSN)

  • 0006-2960

Digital Object Identifier (DOI)

  • 10.1021/bi00037a038

Language

  • eng