Isolation and partial purification of a novel type II restriction endonuclease Bsu121 i, from Bacillus subtilis: Bsu121I, a type II restriction endonuclease from Bacillus subtilis

Abstract

A new type II restriction endonuclease which we designated as Bsu121I has been isolated from gram-positive bacterium Bacillus subtilis strain 121 and partially purified. The restriction endonuclease was isolated from cell extracts using step-wise purification through ammonium sulfate precipitation, followed by phosphocellulose column chromatography. SDS-PAGE profile showed denatured molecular weights (23 and 67 kDa) of the endonuclease. The partially purified enzyme restricted pBR322 DNA into two fragments of 3200 and 1700 bp. The endonuclease activity required Mg+2 as cofactor like other type II endonucleases.