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[Testing and isolation of high-purity restriction endonucleases]
L I Puchkova1, T A Ushakova, V K Mikhaĭlova
1Vector State Research Center for Virology and Biotechnology, Kol'tsovo, Novosibirsk Oblast, 630559 Russia.
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.