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PRELIMINARY CHARACTERIZATION AND PARTIAL-PURIFICATION OF RIBOSOMAL GENE PROMOTER-BINDING PROTEINS FROM TRYPANOSOMA-BRUCEI
journal contribution
posted on 2023-06-07, 23:22 authored by P W Mburu, T J BeebeeDNA 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.
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Publication status
- Published
Journal
BBA - Biochimica et Biophysica ActaISSN
0006-3002Publisher
ElsevierIssue
1-2Volume
1172Page range
5-11Department affiliated with
- Evolution, Behaviour and Environment Publications
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- No
Peer reviewed?
- Yes
Legacy Posted Date
2012-02-06Usage metrics
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