一种来自蜡样芽孢杆菌的Sau3A I限制内切酶同裂酶。

An Sau3 AI restriction endonuclease isoschizomer from Bacillus cereus.

作者信息

Cruz A K, Kidane G, Pires M Q, Rabinovitch L, Guaycurus T V, Morel C M

出版信息

FEBS Lett. 1984 Jul 23;173(1):99-102. doi: 10.1016/0014-5793(84)81025-x.

Abstract

The isolation and characterization of a restriction endonuclease from Bacillus cereus IOC 243 are described. The enzyme recognizes the palindromic sequence 5'-G(met-A,A)TC-3' as determined by PEI chromatography of pancreatic DNase, snake venom phosphodiesterase digestion products of labelled fragments, analysis of restriction digests from normal and N6-methyladenine-free DNA and direct sequence analysis of cloned fragments. The staggered cleavage products with 5' -terminal pGATC extensions are efficiently labelled with polynucleotide kinase and are easily cloned into BamHI sites. The enzyme, denoted Bce243, is thus an isoschizomer of Sau3AI. Its use and potential advantages in substituting Sau3AI. are discussed.

摘要

本文描述了从蜡样芽孢杆菌IOC 243中分离和鉴定一种限制性内切酶的过程。通过对胰腺DNA酶的聚乙二醇离子交换柱层析、标记片段的蛇毒磷酸二酯酶消化产物、正常DNA和无N6 - 甲基腺嘌呤DNA的限制性酶切分析以及克隆片段的直接序列分析，确定该酶识别回文序列5'-G(met - A,A)TC - 3'。具有5'-末端pGATC延伸的交错切割产物可被多核苷酸激酶有效标记，并易于克隆到BamHI位点。这种酶被命名为Bce243，是Sau3AI的同裂酶。文中讨论了其在替代Sau3AI方面的用途和潜在优势。