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Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei
Nucleic Acids Research · 1983 · Vol. 11(5) · pp. 1475–1489
John David Dignam✉(Washington University in St. Louis)Russell M. Lebovitz(Washington University in St. Louis)R G Roeder(Washington University in St. Louis)
Abstract
We have developed a procedure for preparing extracts from nuclei of human tissue culture cells that directs accurate transcription initiation in vitro from class II promoters. Conditions of extraction and assay have been optimized for maximum activity using the major late promoter of adenovirus 2. The extract also directs accurate transcription initiation from other adenovirus promoters and cellular promoters. The extract also directs accurate transcription initiation from class III promoters (tRNA and Ad 2 VA).
Virus-based gene therapy researchViral Infectious Diseases and Gene Expression in InsectsRNA and protein synthesis mechanismsPromoterBiologyTranscription (linguistics)RNA polymerase IIMolecular biologyTranscription factor II DRNA polymeraseGeneral transcription factorRNAGenetics
MeSH terms
AnimalsCattleCell NucleusHeLa CellsHumansKineticsMagnesiumMolecular WeightOperonProtease InhibitorsRNA Polymerase IIDNA-Directed RNA PolymerasesRNA, NeoplasmRNA, TransferTemperature
Funding
- National Institutes of Health
- National Cancer Institute
- National Institute of General Medical Sciences
Citations
11,601
FWCI
41.99
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22
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