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Purification and Subunit Structure of DNA‐dependent RNA Polymerase BII from Rat Brain Nuclei 1
Author(s) -
Yamamoto H.,
Takahashi Y.
Publication year - 1980
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.1980.tb06590.x
Subject(s) - dithiothreitol , protein subunit , enzyme , gel electrophoresis , polyacrylamide gel electrophoresis , microbiology and biotechnology , chemistry , glycerol , chromatography , centrifugation , dna , biochemistry , rna , electrophoresis , polymerase , biology , gene
DNA‐dependent RNA polymerase BII from rat brain nuclei was purified by ammonium sulphate fractionation, successive chromatography on ion‐exchange resins, and glycerol density gradient centrifugation. The purified enzyme was resolved into two components (BIIa and BIIb) on native polyacrylamide gel electrophoresis. The subunit compositions of these enzymes were analysed on SDS‐polyacrylamide gel. BIIa enzyme consisted of nine subunits: BII1 (215,000), BII3 (150,000), BII4 (35,000), BII5 (25,000), BII6 (21,500), BII7 (17,500), BII8 (15,500), BII9 (14,500), and BII10 (11,500); in BIIb enzyme, which also contained nine subunits, BII1 was replaced by a BII2 (180,000) subunit. Addition of both 30% glycerol and dithiothreitol to the preparation medium was essential for stabilizing the enzyme.