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Molecular Biology Reports

, Volume 14, Issue 4, pp 247–249 | Cite as

Inactivation of DNA polymerases by adenosine 2′,3′-riboepoxide 5′-triphosphate allows estimation of the primers affinity

  • Vladimir N. Podust
  • Tamara O. Korobeinicheva
  • George A. Nevinsky
  • Asja S. Levina
  • Olga I. Lavrik
Article

Abstract

Template-primer dependent inactivation of human DNA polymerase α and Klenow fragment of E. coli DNA polymerase I by adenosine 2′,3′-riboepoxide 5′-triphosphate was used for quantitative analysis of the Kd values for oligonucleotide primers of different length. The Kd values are smaller by a factor of 2.5 than the Km values for the same primers determined in the reaction of DNA polymerization in the case of DNA polymerase α. The Kd and Km values are nearly the same for Klenow fragment. Such approach to the determination of Km/Kd ratio can likely be used for detailed quantitative analysis of DNA polymerases.

Key words

DNA polymerase enzyme inactivation primer Kd value 

Abbreviations

epATP

adenosine 2′,3′-riboepoxide 5′-triphosphate

KF

Klenow fragment of E. coli DNA polymerase I

Pol I

E. coli DNA polymerase I

Pol α

human placenta DNA polymerase α

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Copyright information

© Kluwer Academic Publishers 1990

Authors and Affiliations

  • Vladimir N. Podust
    • 1
  • Tamara O. Korobeinicheva
    • 1
  • George A. Nevinsky
    • 1
  • Asja S. Levina
    • 1
  • Olga I. Lavrik
    • 1
  1. 1.Institute of Bioorganic ChemistrySiberian Division of the USSR Academy of SciencesNovosibirskUSSR

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