Isolation of restriction enzyme EcoVIII, an isoschizomer of HindIII, produced by Escherichia coli E1585-68.
A restriction endonuclease designated EcoVIII, an isoschizomer of HindIII, was isolated from Escherichia coli E1585-68 and purified by dextran-polyethylene glycol (DPG) phase partition, ammonium sulfate precipitation, phospho- and DEAE-cellulose chromatography, and hydroxylapatite chromatography. The purified EcoVIII was stable during the purification procedure and its high specific activity required 10 mM Mg2+. Unlike HindIII, EcoVIII exhibited a high specific activity even at low pH (pH 6.3) and showed the highest activity at 48 degrees C. Transformation of purified plasmid DNA from E. coli E1585-68 into K-12 indicated that the EcoVIII gene was carried on a multicopy 4.4-kb miniplasmid. EcoVIII seems to be preferable to HindIII for its production and use because of easier handling of producer cells and a wider range of activity.
D001483:Base Sequence; D004262:DNA Restriction Enzymes / Q000302:isolation & purification*; D015247:Deoxyribonuclease HindIII; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D010957:Plasmids; D013379:Substrate Specificity
D004262:DNA Restriction Enzymes; D015247:Deoxyribonuclease HindIII