Rapid, single-step purification of restriction endonucleases on Cibacron Blue F3GA-agarose

Abstract

After sonication and high-speed centrifugation, crude extracts of Bacillus amyloliquefaciens, Providencia alcalifaciens, Xanthomonas holicola and B. globigii were adsorbed on the dye Cibacron blue F3GA covalently cross-linked to agarose. The restriction endonucleases BamHI, PalI, XhoI and Bg/I together with BglII were isolated by elution of the dye column with linear gradients to 0.5 M NaCl. The enzymes so purified were free of contaminating nucleic acids and other nucleases and were sufficiently concentrated for direct, specific DNA hydrolysis.