Open AccessAnimal DNA‐Dependent RNA Polymerases
Author(s) -
Kedinger Claude,
Chambon Pierre
Publication year - 1972
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1972.tb01912.x
Subject(s) - polymerase , enzyme , dna , glycerol , rna , centrifugation , chemistry , dna polymerase , biochemistry , chromatography , rna polymerase , microbiology and biotechnology , biology , gene
This paper describes a method for the purification of calf thymus DNA‐dependent RNA polymerase B activity and its subsequent resolution into two enzymes, BI and BII. The method of purification of the combined B activities involved chromatography on phosphocellulose and hydroxyapatite and centrifugation through a glycerol density gradient. Enzymes BI and BII were separated by two successive chromatographic steps on DEAE‐cellulose. Addition of 30% glycerol to all buffers increased the stability of the enzymes throughout the purification. The polymerases were over 95% pure and free of contaminating nucleases. It seems unlikely that enzyme BII could derive from enzyme BI by a specific proteolytic process occurring during purification of the B activity.