Literature DB >> 10187910

Site-specific endonuclease NspLKI is an isoschizomer of endonuclease HaeIII.

E V Zabaznaya1, L A Zheleznaya, I V Svad'bina, N I Matvienko.   

Abstract

Site-specific endonuclease NspLKI has been isolated and purified to functionally pure state from soil bacterium Nocardia species LK by successive chromatography on columns with phosphocellulose, HTP hydroxyapatite, and heparin-Sepharose. The isolated enzyme recognizes the 5'-GG downward arrowCC-3' sequence on DNA and cleaves it as indicated by the arrow, i.e., it is an isoschizomer of HaeIII. The final enzyme yield is 1.105 units per gram of wet biomass. The enzyme is active in the temperature range of 25-60 degrees C with an optimum at 48-55 degrees C; it does not lose activity on storage for three days at room temperature. An optimal buffer is HRB containing 10 mM Tris-HCl, pH 7.4, 200 microgram/ml albumin, 10 mM MgCl2, and 100 mM NaCl.

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Year:  1999        PMID: 10187910

Source DB:  PubMed          Journal:  Biochemistry (Mosc)        ISSN: 0006-2979            Impact factor:   2.487


  1 in total

1.  Identification and characterization of CbeI, a novel thermostable restriction enzyme from Caldicellulosiruptor bescii DSM 6725 and a member of a new subfamily of HaeIII-like enzymes.

Authors:  Dae-Hwan Chung; Jennifer R Huddleston; Joel Farkas; Janet Westpheling
Journal:  J Ind Microbiol Biotechnol       Date:  2011-05-22       Impact factor: 3.346

  1 in total

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