Print ISSN: 1681-6900

Online ISSN: 2412-0758

Keywords : restriction enzyme

A Restriction Enzyme from Escherichia coli Purification and General Properties

Mukaram Shikara; Nadia Tariq Barakat; Maysem Modaffer Al-Obaidy

Engineering and Technology Journal, 2009, Volume 27, Issue 5, Pages 954-961

An endonuclease restriction enzyme has been purified from E. coli about 40-fold with
DNAse and RNAse recoveries of about 3%. The purification steps included precipitation
of the enzyme with ammonium sulphate, and reclaimed it through Sephadex G-100 and
DEAE-cellulose chromatography. The purified endonuclease was able to break lambda
DNA into three bands. The enzyme has 5% of carbohydrate moiety which means it is a
glycoprotein. Lastly, the comparison with other commercial restriction endonucleases
proves that this enzyme is a restriction enzyme with enzymic activity dependent on