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Purification of DNA-dependent RNA polymerase II from Saccharomyces cerevisiae

✍ Scribed by Grant A. Bitter

Publisher
Elsevier Science
Year
1983
Tongue
English
Weight
807 KB
Volume
128
Category
Article
ISSN
0003-2697

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✦ Synopsis

A rapid procedure for the purification of RNA polymerase II from Saccharomyces cerevisiae is described. Total RNA polymerase activity was solubilized from whole cells by sonication in 0.32 M (NH4)2SO4 and RNA polymerase II purified by polyethylenimine fractionation, ammonium sulfate precipitation, and chromatography on DEAE-cellulose, DEAE-Sephadex, and phosphocellulose. The procedure may be completed in 2.5 days and the resultant enzyme is judged to be greater than 90% pure.

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## Abstract DNA‐dependent RNA polymerase II has been purified from lymphocytes of patients with chronic lymphocytic leukemia (CLL). Form II polymerase and Escherichia coli RNA polymerase have been used to study the transcription of human DNA and chromatin. The hybridization kinetics of the transcri