PRELIMINARY CHARACTERIZATION AND PARTIAL-PURIFICATION OF RIBOSOMAL GENE PROMOTER-BINDING PROTEINS FROM TRYPANOSOMA-BRUCEI

Mburu, P W and Beebee, T J (1993) PRELIMINARY CHARACTERIZATION AND PARTIAL-PURIFICATION OF RIBOSOMAL GENE PROMOTER-BINDING PROTEINS FROM TRYPANOSOMA-BRUCEI. BBA - Biochimica et Biophysica Acta, 1172 (1-2). pp. 5-11. ISSN 0006-3002

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Abstract

DNA fragments including the promoter region of the major ribosomal RNA gene of Trypanosoma brucei (r-promoter) were identified and subcloned using a synthetic oligonucleotide probe corresponding to the putative core promoter. These fragments were used in mobility shift assays with proteins extracted from T brucei nuclei, and demonstrated the presence in 0.15 M NaCl extracts of protein(s) with specific binding affinities for the r-promoter region. Binding was stable in the presence of a 100-fold excess of competitor DNA, and occurred at the relatively low salt concentrations (< 50 mM NaCl) characteristic of many enzyme activity optima in this organism. A control DNA fragment not including the r-promoter region was not retarded in the mobility shift assays, and the r-promoter-binding activity had a molecular weight of about 140 000. Nuclear extracts from T. brucei contained large amounts of DNase activity, and the promoter-binding proteins were partially purified from the crude extract using ammonium sulphate precipitation, sephacryl S-200 and Heparin-sepharose chromatography.

Item Type: Article
Schools and Departments: School of Life Sciences > Evolution, Behaviour and Environment
Depositing User: Trevor Beebee
Date Deposited: 06 Feb 2012 19:33
Last Modified: 08 Jun 2012 14:01
URI: http://sro.sussex.ac.uk/id/eprint/21264
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