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An express method for testing the activity of a repair enzyme, uracil-DNA-glycosylase

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Abstract

A rapid and effective method for testing a repair enzyme, uracil-DNA-glycosylase, was proposed. As a substrate, a deoxyuridine-containing 5′-32P-labeled deoxyoligonucleotide covalently attached to a polystyrene support (Tenta Gel S-NH2) was used. The ammonia cleavage of the apyrimidine site formed in the enzymic reaction followed by the transition of the labeled oligonucleotide fragment from the solid phase into solution allowed the detection of the enzymic activity.

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Abbreviations

AP site:

apyrimidine/apurine site

[3H]dU-DNA:

oligonucleotide containing [3H]-deoxyuridine

UDG:

uracil-DNA-glycosylase

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Authors and Affiliations

  1. Chemical Faculty, Moscow State University, Vorob’evy gory, 119899, GSP-3, Moscow, Russia

    A. E. Sud’ina, E. M. Volkov & T. S. Oretskaya

  2. Institute of Molecular Biology and Biophysics, Siberian Division, Russian Academy of Medical Sciences, Novosibirsk, Russia

    S. Kh. Degtyarev & D. A. Gonchar

  3. Belozerskii Institute of Physicochemical Biology, Moscow State University, Moscow, Russia

    E. A. Kubareva

Authors
  1. A. E. Sud’ina
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  2. E. M. Volkov
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  3. T. S. Oretskaya
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  4. S. Kh. Degtyarev
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  5. D. A. Gonchar
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  6. E. A. Kubareva
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Correspondence to E. A. Kubareva.

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Sud’ina, A.E., Volkov, E.M., Oretskaya, T.S. et al. An express method for testing the activity of a repair enzyme, uracil-DNA-glycosylase. Russ J Bioorg Chem 26, 398–402 (2000). https://doi.org/10.1007/BF02758667

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  • DOI: https://doi.org/10.1007/BF02758667

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