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来自沙门氏菌属的两种I型限制酶。纯化及DNA识别序列。

Two type I restriction enzymes from Salmonella species. Purification and DNA recognition sequences.

作者信息

Nagaraja V, Shepherd J C, Pripfl T, Bickle T A

出版信息

J Mol Biol. 1985 Apr 20;182(4):579-87. doi: 10.1016/0022-2836(85)90243-8.

Abstract

We have purified the type I restriction enzymes SB and SP from Salmonella typhimurium and S. potsdam, respectively, and determined the DNA sequences that they recognize. These sequences resemble those previously determined for the type I enzymes, EcoB, EcoK and EcoA, in that the specific part of the sequence is divided into two domains by a spacer of non-specific sequence that has a fixed length for each enzyme. Two main differences from the previously determined sequences are seen. Both of the new sequences are degenerate and one of them, SB, has one trinucleotide and one pentanucleotide-specific domain rather than the trinucleotide and tetranucleotide domains seen for all of the other enzymes. The only conserved features of the recognition sequences are the adenosyl residues that are methylated in the modification reaction. For all of the enzymes these are situated ten or 11 base-pairs apart, one on each strand of the DNA. This suggests that the enzymes bind to DNA along one face of the double helix making protein-DNA interaction in two successive major grooves with most of the non-specific spacer sequence in the intervening minor groove.

摘要

我们分别从鼠伤寒沙门氏菌和波茨坦沙门氏菌中纯化出了I型限制酶SB和SP,并确定了它们识别的DNA序列。这些序列与先前确定的I型酶EcoB、EcoK和EcoA的序列相似,即序列的特定部分被一段非特异性序列间隔区分为两个结构域,每个酶的间隔区长度固定。与先前确定的序列有两个主要差异。这两个新序列都是简并的,其中一个序列SB有一个三核苷酸和一个五核苷酸特异性结构域,而不是其他所有酶都有的三核苷酸和四核苷酸结构域。识别序列中唯一保守的特征是在修饰反应中被甲基化的腺苷残基。对于所有这些酶来说,这些残基相隔10或11个碱基对,位于DNA的每条链上一个。这表明这些酶沿着双螺旋的一个面与DNA结合,在两个连续的大沟中形成蛋白质-DNA相互作用,中间的小沟中有大部分非特异性间隔序列。

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