来自芽孢杆菌属菌株d6的位点特异性切口酶

Site-specific nickase from bacillus species strain d6.

作者信息

Zheleznaya L A, Perevyazova T A, Alzhanova D V, Matvienko N I

机构信息

Institute of Theoretical and Experimental Biophysics, Russian Academy of Sciences, Pushchino, Moscow Region, 142290 Russia.

出版信息

Biochemistry (Mosc). 2001 Sep;66(9):989-93. doi: 10.1023/a:1012369525809.

DOI:10.1023/a:1012369525809

PMID:11703181

Abstract

Three site-specific endonucleases were found in thermophilic strain Bacillus species D6. One of them, BspD6II, is an isoschizomer of Eco57I. The second, BspD6III, is present in the strain in very small amount; therefore, it has not been characterized. This paper is devoted to the third, BspD6I, which recognizes pentanucleotide site 5'-GAGTC-3' and cleaves only one DNA strand at a distance of 4 nucleotides from the site in the 3'-direction in the chain with the GAGTC sequence, i.e., it behaves as a site-specific nickase. Nickase N.BspD6I cleaves one DNA strand only in double-stranded DNA and does not cleave single-stranded DNA. Site-specific methylase SscL1I (an isohypectomer of M*HinfI) that methylates adenine in the sequence 5'-GAGTC-3' prevents DNA hydrolysis by nickase BspD6I.

摘要

在嗜热菌株芽孢杆菌属D6中发现了三种位点特异性内切核酸酶。其中一种，BspD6II，是Eco57I的同裂酶。第二种，BspD6III，在该菌株中的含量非常少；因此，尚未对其进行表征。本文致力于研究第三种酶，BspD6I，它识别五核苷酸位点5'-GAGTC-3'，并仅在具有GAGTC序列的链中从该位点沿3'方向4个核苷酸处切割一条DNA链，即它表现为位点特异性切口酶。切口酶N.BspD6I仅在双链DNA中切割一条DNA链，而不切割单链DNA。在序列5'-GAGTC-3'中使腺嘌呤甲基化的位点特异性甲基化酶SscL1I（M*HinfI的同型异构体）可防止切口酶BspD6I水解DNA。