title: [Testing and isolation of high-purity restriction endonucleases].
abstract: A new method of testing restriction nucleases is proposed. This method is based on high-temperature treatment of crude cell extracts. Disrupted cells were heated at 50-60 degrees C, centrifuged, and assayed for restrictases. This method provides the opportunity for screening new enzymes in microbial strains enriched with nonspecific restrictases. High-temperature treatment of cell extracts of certain producers reduces the number of steps of the procedure used for isolating high-purity restrictases; the resulting preparations are capable of maintaining high enzymatic activity during long-term storage. It was shown that high-temperature treatment can be applied not only to thermophilic but also to mesophilic strains of microorganisms of different taxa.
authors.affiliation: Vector State Research Center for Virology and Biotechnology, Kol'tsovo, Novosibirsk Oblast, 630559 Russia.
authors.forename: L I
authors.initials: LI
authors.lastname: Puchkova
authors.forename: T A
authors.initials: TA
authors.lastname: Ushakova
authors.forename: V K
authors.initials: VK
authors.lastname: Mikhaĭlova
authors.forename: G D
authors.initials: GD
authors.lastname: Serov
authors.forename: G N
authors.initials: GN
authors.lastname: Krivopalova
authors.forename: V E
authors.initials: VE
authors.lastname: Repin
mesh_terms: D001419:Bacteria / Q000201:enzymology*; D001709:Biotechnology / Q000379:methods; D004262:DNA Restriction Enzymes / Q000302:isolation & purification*; D006358:Hot Temperature; D011786:Quality Control
chemical_list: D004262:DNA Restriction Enzymes