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

AuthorsBabkina, Olga V.; Chutko, Catherine A.; Shashkov, Alexander A.; Dzhidzhoev, Murat S.; Eritja Casadellà, Ramón ; Gromova, Elizaveta S.
double stranded DNA
Base Sequence
Issue Date1-Sep-2002
PublisherRoyal Society of Chemistry (UK)
CitationPhotochemical and Photobiological Sciences 1 (9): 636-640 (2002)
AbstractWe used a XeCl excimer laser with 50 ns pulses, a frequency of 0.3 Hz and a wavelength of 308 mn 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.
Publisher version (URL)http://dx.doi.org/10.1039/b202028a
Appears in Collections:(IQAC) Artículos
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