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Sequence-specific DNA affinity chromatography: Application to the purification of EcoRI and SphI

1991; Elsevier BV; Volume: 195; Issue: 2 Linguagem: Inglês

10.1016/0003-2697(91)90340-y

1096-0309

G. Vlatakis, Vassilis Bouriotis,

DNA and Nucleic Acid Chemistry

Several rapid and effective methods have been described to obtain restriction endonucleases suitable for commercial exploitation. However, lengthy and laborious protocols have been necessary to obtain homogeneous enzymes. We now report the use of sequence-specific DNA affinity chromatography to purify restriction endonucleases to near homogeneity. Restriction endonucleases EcoRI and SphI from the micro-organisms Escherichia coli RY 13 and Streptomyces phaeochromogenes, respectively, were purified to near homogeneity employing a two-step procedure which involves DNA-cellulose chromatography and oligonucleotide-ligand affinity chromatography.