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Cloning, expression and functional characterization of Schizosaccharomyces pombe TFIIB
Evelyn Tamayo1, Edio Maldonado
1Programa de Biologia Celular y Molecular, Facultad de Medicina, Instituto de Ciencias Biomedicas, Universidad de Chile, Independencia 1027, Casilla 70086, Santiago 7, Chile.
Abstract:
The transcription factor TFIIB has been identified and cloned from the yeast Schizosaccharomyces pombe. The cloned polypeptide is highly homologous to human TFIIB and to Saccharomyces cerevisiae TFIIB. S. pombe TFIIB is a 340-amino-acid-long protein and it possesses a repeated motif of 75 amino acids near the carboxy-terminal region. The purified recombinant protein is able to bind to the TBP-DNA promoter complex in gel retardation experiments. Recombinant S. pombe TFIIB is active in in vitro transcription assays, since it can complement the transcription activity of a S. pombe cell extract in which TFIIB was depleted by using antibodies.