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

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.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Chromatography, DEAE-Cellulose
  • Chromatography, Ion Exchange
  • DNA-Directed RNA Polymerases / isolation & purification*
  • Drug Stability
  • Electrophoresis, Polyacrylamide Gel
  • Polyethyleneimine
  • Triticum / enzymology*

Substances

  • Polyethyleneimine
  • DNA-Directed RNA Polymerases