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Iodouracil-mediated photocrosslinking of DNA to EcoRII restriction endonuclease in catalytic conditions

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Abstract

We used a XeCl excimer laser with 50 ns pulses, a frequency of 0.3 Hz and a wavelength of 308 nm in appropriate conditions for the photocrosslinking of EcoRII restriction endonuclease to a 14-mer DNA duplex, containing a 5-iodo-2’-deoxyuridine residue (IdU). IdU replaced the thymidine residue within the EcoRII recognition sequence 5’-CCT/AGG. The binding of EcoRII endonuclease to the IdU-containing DNA duplex was analyzed by gel retardation assay in the presence of Ca2+ or Mg2+ ions. Photocrosslinking of EcoRII to the IdU-containing DNA duplex occurred in a pre-reactive complex formed in the presence of Ca2+ ions. Photocrosslinking yields as a function of time and UV-laser light intensity were studied.

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Authors and Affiliations

  1. Department of Chemistry and Belozersky Institute of Physical and Chemical Biology, Moscow State University, Moscow, 119992, Russia

    Olga V. Babkina & Elizaveta S. Gromova

  2. Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, 117984, Russia

    Olga V. Babkina

  3. Department of Physics, Moscow State University, Moscow, 119992, Russia

    Catherine A. Chutko, Alexander A. Shashkov & Murat S. Dzhidzhoev

  4. Centro de Investigación y Desarrollo, C.S.I.C., Jordi Girona 18-26, 08034, Barcelona, Spain

    Ramon Eritja

Authors
  1. Olga V. Babkina
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  2. Catherine A. Chutko
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  3. Alexander A. Shashkov
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  4. Murat S. Dzhidzhoev
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  5. Ramon Eritja
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  6. Elizaveta S. Gromova
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Correspondence to Elizaveta S. Gromova.

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Babkina, O.V., Chutko, C.A., Shashkov, A.A. et al. Iodouracil-mediated photocrosslinking of DNA to EcoRII restriction endonuclease in catalytic conditions. Photochem Photobiol Sci 1, 636–640 (2002). https://doi.org/10.1039/b202028a

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  • DOI: https://doi.org/10.1039/b202028a

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