[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 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.
Puchkova
L I
LI
Vector State Research Center for Virology and Biotechnology, Kol'tsovo, Novosibirsk Oblast, 630559 Russia.
Ushakova
T A
TA
Mikhaĭlova
V K
VK
Serov
G D
GD
Krivopalova
G N
GN
Repin
V E
VE
D001419:Bacteria / Q000201:enzymology*; D001709:Biotechnology / Q000379:methods; D004262:DNA Restriction Enzymes / Q000302:isolation & purification*; D006358:Hot Temperature; D011786:Quality Control
D004262:DNA Restriction Enzymes