Cloning and sequencing of the gene of site-specific nickase N.BspD6I

A fragment of chromosomal DNA from Bacillus species D6 containing the gene of nickase N.BspD6I and the regions adjacent to its 5;- and 3;-ends was cloned and sequenced. The nucleotide sequence of the nickase gene, except of one neutral change, is homologous to the nicking endonuclease N.BstNBI gene...

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Bibliographic Details
Published in:Biochemistry (Moscow) 2003-09, Vol.68 (9), p.984-987
Main Authors: Perevyazova, T A, Rogulin, E A, Zheleznaya, L A, Matvienko, N I
Format: Article
Language:English
Subjects:
Bacillus - enzymology
Bacillus - genetics
Bacteria
Bacteriology
Cloning
Cloning, Molecular
Deoxyribonuclease I - genetics
DNA Restriction Enzymes - genetics
DNA, Bacterial - genetics
E coli
Molecular Sequence Data
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Summary:A fragment of chromosomal DNA from Bacillus species D6 containing the gene of nickase N.BspD6I and the regions adjacent to its 5;- and 3;-ends was cloned and sequenced. The nucleotide sequence of the nickase gene, except of one neutral change, is homologous to the nicking endonuclease N.BstNBI gene sequenced by Higgens et al. (2001). After integration of a PCR-copy of the nickase gene into an expression vector pET28b under the control of the phage T7 promoter, specific nicking activity was detected in the lysates of transformed E. coli cells.
ISSN:0006-2979
1608-3040
DOI:10.1023/A:1026008528310