来自天蓝色链霉菌的一种核酸内切酶（E.C. 3.1.30.1）的纯化与特性分析

Purification and characterization of an endonuclease (E.C. 3.1.30.1) from Streptomyces tendae.

作者信息

Engel P, Ullah A H

机构信息

Southern Regional Research Laboratory, ARS, New Orleans, Louisiana 70124.

出版信息

Prep Biochem. 1988;18(2):137-52. doi: 10.1080/00327488808062517.

DOI:10.1080/00327488808062517

PMID:2836846

Abstract

A single-strand-specific endonuclease which converted negatively supercoiled DNA to open-circular and linear DNA was purified to homogeneity with Hb-Sepharose 4B, DEAE Trisacryl M, HA-Ultrogel and PBE-94 chromatofocusing from extracts of Streptomyces tendae ATCC 31160. Bio-Gel P-200 chromatography and electrophoresis in SDS-PAGE indicated the native protein was a monomer with a molecular weight of approximately 40-kDa. This enzyme did not hydrolyze double-stranded linear DNA but digested RNA and circular single-strand DNA. Sequence specificity for nicking of negatively supercoiled DNA was not detected.

摘要

从天蓝色链霉菌ATCC 31160提取物中，利用Hb-Sepharose 4B、DEAE Trisacryl M、HA-Ultrogel和PBE-94色谱聚焦法，纯化得到一种单链特异性核酸内切酶，该酶可将负超螺旋DNA转化为开环和线性DNA。Bio-Gel P-200色谱法和SDS-PAGE电泳表明，天然蛋白是一种分子量约为40 kDa的单体。该酶不水解双链线性DNA，但能消化RNA和环状单链DNA。未检测到负超螺旋DNA切口的序列特异性。