Assays for investigating transcription by RNA polymerase II in vitro.
With the availability of the general initiation factors (TFIIB, TFIID, TFIIE, TFIIF, and TFIIH), it is now possible to investigate aspects of the mechanism of eukaryotic messenger RNA synthesis in purified, reconstituted RNA polymerase II transcription systems. Rapid progress in these investigations has been spurred by use of a growing number of assays that are proving valuable not only for dissecting the molecular mechanisms of transcription initiation and elongation by RNA polymerase II, but also for identifying and purifying novel transcription factors that regulate polymerase activity. Here we describe a variety of these assays and discuss their utility in the analysis of transcription by RNA polymerase II.
Reines
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Department of Biochemistry, Emory University School of Medicine, Atlanta, Georgia 30322, USA.
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Conaway
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Conaway
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D002474:Cell-Free System / Q000201:enzymology; D005821:Genetic Techniques; D012319:RNA Polymerase II / Q000235:genetics*; D014158:Transcription, Genetic
D012319:RNA Polymerase II