Rapid, single-step purification of restriction endonucleases on cibacron blue F3GA-agarose.
After sonication and high-speed centrifugation, crude extracts of B. amyloliquefaciens, P. alcalifaciens, X. holicola, and B. globiggi were adsorbed on the dye Cibacron blue F3GA convalently cross-linked to agarose. The restriction endonucleases BamHI, PalI, XhoI, and BglI 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.
D000873:Anthracenes; D001407:Bacillus / Q000201:enzymology; D001419:Bacteria / Q000201:enzymology; D002846:Chromatography, Affinity; D004396:Coloring Agents; D004262:DNA Restriction Enzymes / Q000302:isolation & purification*; D011549:Pseudomonas / Q000201:enzymology; D012685:Sepharose; D013045:Species Specificity; D014227:Triazines
D000873:Anthracenes; D004396:Coloring Agents; D014227:Triazines; C012842:Cibacron Blue F 3GA; D012685:Sepharose; D004262:DNA Restriction Enzymes