A new method for the large-scale purification of wheat germ DNA-dependent RNA polymerase II.
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.
D002848:Chromatography, DEAE-Cellulose; D002852:Chromatography, Ion Exchange; D012321:DNA-Directed RNA Polymerases / Q000302:isolation & purification*; D004355:Drug Stability; D004591:Electrophoresis, Polyacrylamide Gel; D011094:Polyethyleneimine; D014908:Triticum / Q000201:enzymology*
D011094:Polyethyleneimine; D012321:DNA-Directed RNA Polymerases