SEPARATION OF ESCHERICHIA-COLI RNA-POLYMERASE SIGMA-70 HOLOENZYME FROM CORE ENZYME ON HEPARIN-SEPHAROSE COLUMNS

被引:5
作者
WELLINGTON, SR [1 ]
SPIEGELMAN, GB [1 ]
机构
[1] UNIV BRITISH COLUMBIA,DEPT MICROBIOL,VANCOUVER V6T 1W5,BC,CANADA
基金
加拿大自然科学与工程研究理事会;
关键词
D O I
10.1016/0006-291X(91)91934-5
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
A method is described for the rapid purification of DNA-dependent RNA polymerase sigma-70 holoenzyme from Escherichia coli. The essential step in this protocol involves the differential elution of sigma-70 holoenzyme from core polymerase on a heparin-Sepharose column. Using a linear gradient of KCl, holoenzyme was found to elute at 0.25M whereas core polymerase eluted at 0.35 M. From 20 g of cells, up to 1 mg of RNA polymerase holoenzyme could be isolated in two days. The preparations were greater than 95% pure with respect to protein, and saturated with the sigma subunit. © 1991.
引用
收藏
页码:1107 / 1114
页数:8
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