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Isolation and some properties of restriction endonuclease from Bacillus amyloliquefaciens

Sokolov, N.N.; Votrin, I.I.; Fitzner, A.B.; Anikejcheva, N.V.

Biokhimiia 43(5): 865-871

1978

ISSN/ISBN: 0320-9725
PMID: 656508
Document Number: 137517
A partially purified preparation of restriction endonuclease Bam I was isolated from the cells of Bacillus amyloliquefaciens. The purification procedure was a modification of a method described by Wilson and Young. Isolation and purification of the enzyme involved disruption of the cells by ultrasonication, treatment with streptomycin sulfate, fractionation by ammonium sulfate, chromatography on DEAE-cellulose and hydroxylapatite and rechromatography on DEAE-cellulose. Restrictase Bam I splits the linear double-chain DNA molecule of phage gamma into six fragments. The enzyme retained its stability under storage on the ice for 1,5--2 months in a Na- or K-phosphate buffer with beta-mercaptoethanol (10 mM).

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