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Eco1524I, a type II restriction endonuclease

Isolation, partial purification, and characterization

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Abstract

Various strains of Escherichia coli, isolated from different patients, were screened for type II restriction endonuclease activity. In 1 out of 23 patients, a type II restriction endonuclease activity was found. The restriction endonuclease designated Eco1524I was purified to near homogeneity, based on hydroxyapatite and heparin sepharose chromatography. Eco1524I exhibited endonuclease restriction activity in the pH range from 6.0 to 10.0 (maximum level at pH 8.0) and required Mg2+ as divalent cation. The enzyme was stable till temperature 55°C and pH range from 6.0 to 10.0. Eco1524I recognized the sequence 6-bp palindromic 5′AGG↑CCT 3′, producing blunt end and is found to be an isoschizomer of Stu I.

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Authors and Affiliations

  1. Laboratoire de Biochimie, Faculté des Sciences de Tunis, Campus Universitaire 1060, Tunis, Tunisia

    Hadeer Lazim & Omrane Belhadj

  2. Department of Food Sciences, Royal Veterinary and Agricultural University, D-1958, Frederiksberg C, Denmark

    Jytte Josephsen

  3. Laboratoire de Microbiologie, Centre National de Greffe de Moelle osseuse, Bab Saadoun Tunis, Tunisia

    Assia Ben Hassen

  4. Laboratoire Interactions Légumineuses-Microorganismes, Institut National de Recherche Scientifique et Technique, 95-2050, Hammam-Lif, Tunisia

    Férid Limam

Authors
  1. Hadeer Lazim
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  2. Jytte Josephsen
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  3. Assia Ben Hassen
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  4. Omrane Belhadj
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  5. Férid Limam
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Correspondence to Férid Limam.

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Lazim, H., Josephsen, J., Hassen, A.B. et al. Eco1524I, a type II restriction endonuclease. Appl Biochem Biotechnol 125, 189–199 (2005). https://doi.org/10.1385/ABAB:125:3:189

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  • DOI: https://doi.org/10.1385/ABAB:125:3:189

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