Site-specific endonuclease NspLKI is an isoschizomer of endonuclease HaeIII.
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.
Zabaznaya
E V
EV
Institute of Protein Research, Russian Academy of Sciences, Pushchino, Moscow Region, 142292, Russia. nikmatv@vega.protres.ru
Zheleznaya
L A
LA
Svad'bina
I V
IV
Matvienko
N I
NI
D002852:Chromatography, Ion Exchange; D004247:DNA / Q000378:metabolism; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism*; D004587:Electrophoresis, Agar Gel; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D013379:Substrate Specificity; D013696:Temperature
D004247:DNA; D015252:Deoxyribonucleases, Type II Site-Specific; C024403:GGCC-specific type II deoxyribonucleases