Purification of calf thymus RNA polymerase II for in vitro transcription studies.
A procedure for the purification of a large amount of RNA polymerase II (RNAP II) from calf thymus is described. This procedure results in an approximately 1400-fold purification with 40% yield of enzyme in 60 h. The partially purified enzyme is highly suitable for in vitro transcription studies in a cell-free system utilizing HeLa S-100. This method for RNAP II purification would find significant applications in in vitro studies for the analysis of factors modulating eukaryotic transcription.
Tyagi
J S
JS
Laboratory of Molecular Biology, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892.
Pastan
I
I
D000818:Animals; D002417:Cattle; D002474:Cell-Free System; D002642:Chick Embryo; D004795:Enzyme Stability; D012319:RNA Polymerase II / Q000302:isolation & purification*; D013698:Templates, Genetic; D013950:Thymus Gland / Q000201:enzymology*; D014158:Transcription, Genetic
D012319:RNA Polymerase II