A new method for the purification of RNA polymerase II (or B) from the lower eukaryote Physarum polycephalum. The presence of subforms.
The DNA-dependent RNA polymerase II or B from the lower eukaryote Physarum polycephalum has been purified to apparent homogeneity by a new method employing poly(ethylene imine) precipitation and elution, and heparin-Sepharose affinity chromatography. The method is readily scaled up or down and affords a purification of over 5000-fold with a yield of 35-45%. The procedure is easy to perform and can be carried out in less than three days even on a large scale. Furthermore, it gives enzyme of higher purity and in at least 10-fold greater yield than previously published procedures for its purification from this organism. These improvements have allowed the detection of a series of subforms of the enzyme. The combination of precipitation using poly(ethylene imine) with chromatography on heparin-Sepharose may prove useful in the preparation of other proteins which interact with nucleic acids.
D002846:Chromatography, Affinity; D012321:DNA-Directed RNA Polymerases / Q000302:isolation & purification*; D007700:Kinetics; D046911:Macromolecular Substances; D008970:Molecular Weight; D009235:Myxomycetes / Q000201:enzymology*; D009994:Osmolar Concentration; D010804:Physarum / Q000201:enzymology*; D012319:RNA Polymerase II / Q000302:isolation & purification* / Q000378:metabolism
D046911:Macromolecular Substances; D012319:RNA Polymerase II; D012321:DNA-Directed RNA Polymerases