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[嗜热土壤细菌嗜热脂肪芽孢杆菌BS中BstBSI限制性内切酶的分离与特性]

[Isolation and properties of BstBSI restriction endonuclease from the thermophilic soil bacteria Bacillus stearothermophilus BS].

作者信息

Kovalevskaia N P, Ivanov L Iu, Zheleznaia L A, Matvienko N I

出版信息

Mol Gen Mikrobiol Virusol. 1993 May-Jun(3):22-5.

PMID:8350878
Abstract

A new restriction endonuclease BstBSI was isolated and purified from the thermophilic soil bacterium Bacillus stearothermophilus BS by the blue sepharose and hydroxyapatite chromatographies. The enzyme is an isoschizomer of SnaI from Sphaerotilus natans C. It recognizes the hexanucleotide GTATAC and cleaves DNA in the center of the sequence. The maximal catalytic activity of the endonuclease is registered in 50 mM tris-HCl (pH 9.0) buffer with the high ionic strength (100 mM NaCl) in the presence of 10 mM MgCl2 at 45 degrees C. Glucosylated DNA of the phage T4 is not cleaved by the enzyme.

摘要

通过蓝色琼脂糖和羟基磷灰石色谱法从嗜热土壤细菌嗜热脂肪芽孢杆菌BS中分离并纯化出一种新的限制性内切酶BstBSI。该酶是来自浮游球衣菌C的SnaI的同裂酶。它识别六核苷酸GTATAC并在序列中心切割DNA。内切酶的最大催化活性在含有10 mM MgCl2、高离子强度(100 mM NaCl)的50 mM Tris-HCl(pH 9.0)缓冲液中于45℃时测得。噬菌体T4的糖基化DNA不能被该酶切割。

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