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Updated: Aug 9, 2026

High-throughput Purification of Affinity-tagged Recombinant Proteins
Published on: August 26, 2012
Class II (B) general transcription factor (TFIIB) that binds to the template-committed preinitiation complex is
V Moncollin1, L Fischer, B Cavallini
1Centre National de la Recherche Scientifique Unité 184, l'Institut National de la Santé et de la Recherche Médicale, Faculté de Médecine, Strasbourg, France.
Abstract:
A class II (B) general transcription factor of 34 kDa has been purified from HeLa cells to apparent homogeneity. This factor appears to be transcription factor IIB (TFIIB), since it binds in vitro to template-committed preinitiation complexes formed between a template containing the TATA box/cap-site elements of the adenovirus type 2 major late promoter (Ad2MLP) and recombinant human or yeast TFIID (previously called BTF1) expressed in Escherichia coli. DNase I footprint studies show an extended pattern of protection of Ad2MLP TATA box/cap-site sequences when TFIIB is bound to template-committed complexes, even though TFIIB does not bind on its own to the template in the absence of TFIID. We also show that TFIIB is different from BTF3 by a number of criteria.
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