一种大规模纯化小麦胚DNA依赖性RNA聚合酶II的新方法。

A new method for the large-scale purification of wheat germ DNA-dependent RNA polymerase II.

作者信息

Jendrisak J J, Burgess R R

出版信息

Biochemistry. 1975 Oct 21;14(21):4639-45. doi: 10.1021/bi00692a012.

Abstract

An improved method for the purification of the alpha-amanitin-sensitive deoxyribonucleic acid dependent ribonucleic acid polymerase [ribonucleosidetriphosphate:RNA-nucleotidyltransferase, EC 2.7.7.6-A1 (RNA polymerase II or RNA polymerase B) from wheat germ is presented. The method involves homogenization of wheat germ in a buffer of moderate ionic strength, precipitation of RNA polymerase with Polymin P (a polyethylenimine), elution of RNA polymerase from the Polymin P precipitate, ammonium sulfate precipitation, and chromatography on DEAE-cellulose and phosphocellulose. RNA polymerase II is purified over 4000-fold with a 60% recovery, resulting in a yield of 25-30 mg of RNA polymerase from 1 kg of starting material.

摘要

本文介绍了一种改进的从小麦胚芽中纯化对α-鹅膏蕈碱敏感的依赖脱氧核糖核酸的核糖核酸聚合酶[核糖核苷三磷酸：RNA-核苷酸转移酶，EC 2.7.7.6-A1（RNA聚合酶II或RNA聚合酶B）]的方法。该方法包括在中等离子强度的缓冲液中对小麦胚芽进行匀浆，用聚凝胺P（一种聚乙烯亚胺）沉淀RNA聚合酶，从聚凝胺P沉淀中洗脱RNA聚合酶，硫酸铵沉淀，以及在DEAE-纤维素和磷酸纤维素上进行层析。RNA聚合酶II的纯化倍数超过4000倍，回收率为60%，从1千克起始原料中可得到25 - 30毫克RNA聚合酶。