Large-scale purification of wheat germ DNA-dependent RNA polymerase II

作者：Jerome J. Jendrisak, Richard R. Burgess · 发表于：Biochemistry · 年份：1975 · DOI：10.1021/bi00692a012 · 被引用次数：276 · 研究领域：Molecular Biology Techniques and Applications、Plant Disease Resistance and Genetics、Plant Pathogenic Bacteria Studies

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.