A procedure for the rapid, large-scall purification of Escherichia coli DNA-dependent RNA polymerase involving Polymin P precipitation and DNA-cellulose chromatography.
A procedure for the rapid, large-scall purification of Escherichia coli DNA-dependent RNA polymerase involving Polymin P precipitation and DNA-cellulose chromatography.
复制标题
一种快速、大规模纯化大肠杆菌 DNA 依赖性 RNA 聚合酶的方法，涉及 Polymin P 沉淀和 DNA-纤维素色谱。
作者：
R. Burgess;J. Jendrisak
描述了一种从大肠杆菌纯化DNA依赖性RNA聚合酶[三磷酸核糖核苷：RNA核苷酸转移酶，EC 2.7.7.6]的改进方法。该方法包括溶菌酶-脱氧胆酸钠裂解，低速离心，用Polymin P沉淀，从Polymin P沉淀中洗脱，硫酸铵沉淀，以及在DNA-纤维素和Bio-Gel A 5 m上的色谱。RNA聚合酶在2天内纯化至电泳均一，回收率为45%，从500 g细胞中得到250 mg全酶。
An improved method is described for the purification of the DNA-dependent RNA polymerase [ribonucleosidetriphosphate:RNA nucleotidyltransferase, EC 2.7.7.6] from Escherichia coli. The method involves lysozyme-sodium deoxycholate lysis, low-speed centrifugation, precipitation with Polymin P, elution from the Polymin P precipitate, ammonium sulfate precipitation, and chromatography on DNA-cellulose and Bio-Gel A 5m. RNA polymerase is purified to electrophoretic homogeneity in 2 days with a recovery of 45%, resulting in a yield of 250 mg of holoenzyme from 500 g of cells.