R.D. Morgan, C. Calvet, M. Demeter, R. Agra, H. Kong
Characterization of the Specific DNA Nicking Activity of Restriction Endonuclease N.BstNBI
N.BstNBI is a unique restriction endonuclease isolated
from Bacillus stearothermophilus. We have characterized
the recognition sequence and the cleavage
site of N.BstNBI. Mapping of cleavage sites of
N.BstNBI showed that it recognizes an asymmetric
sequence, 5? GAGTC 3?, and cleaves only on the top
strand 4 base pairs away from its recognition sequence.
To verify the nicking activity of N.BstNBI, we
have constructed two plasmids containing a single
recognition sequence (pNB1) or no recognition site
(pNB0). When pNB1 and pNB0 were incubated with
the enzyme, N.BstNBI nicked only the plasmid pNB1,
suggesting that N.BstNBI is a specific nicking endonuclease.
Biological Chemistry, Walter de Gruyter
Print ISSN: 1431-6730
Volume: 381, 11/2000
Pages: 1123 - 1125
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