Testing and Isolation of High-Purity Restriction Endonucleases

Testing and Isolation of High-Purity Restriction Endonucleases

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°C, centrifuged, and assayed for restrictases. This method provides the opportunity for screening new enzymes in microbial strains...

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Veröffentlicht in:Applied biochemistry and microbiology 2002-01, Vol.38 (1), p.15-19
Hauptverfasser: Puchkova, L I, Ushakova, T A, Mikhailova, V K, Serov, G D, Krivopalova, G N, Repin, V E
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Bacteria
Enzymatic activity
High temperature
Microorganisms
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container_title Applied biochemistry and microbiology
container_volume 38
creator Puchkova, L I
Ushakova, T A
Mikhailova, V K
Serov, G D
Krivopalova, G N
Repin, V E
description 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°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.[PUBLICATION ABSTRACT]
doi_str_mv 10.1023/A:1013236218350
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subjects Bacteria
Enzymatic activity
High temperature
Microorganisms
title Testing and Isolation of High-Purity Restriction Endonucleases
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