Partial purification and characterization of RalF40I, a class II restriction endonuclease from Ruminococcus albus F-40, which recognizes and cleaves 5'-/GATC-3'

Restriction endonuclease Ral F40I was purified from cell-free extracts of the rumen cellulolytic bacterium Ruminococcus albus F-40 by heparin-Sepharose chromatography. The preparation was active only on DNA substrates that were not Dam-methylated. Ral F40I recognizes the 4-bp palindrome, 5′-/GATC-3′, and cleaves DNA at the 5′ side of G in the sequence, producing 5′ tetranucleotide protruding ends. Ral F40I is a class II restriction endonuclease and an isoschizomer of Mbo I and Dpn II.