A procedure for the rapid, large-scall purification of Escherichia coli DNA-dependent RNA polymerase involving Polymin P precipitation and DNA-cellulose chromatography.
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.
D002845:Chromatography; D002850:Chromatography, Gel; D012321:DNA-Directed RNA Polymerases / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000201:enzymology*; D011094:Polyethyleneimine
D011094:Polyethyleneimine; D012321:DNA-Directed RNA Polymerases