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Interaction of GAL4 and GAL80 gene regulatory proteins in vitro.

The GAL80 protein of Saccharomyces cerevisiae, synthesized in vitro, bound tightly to GAL4 protein and to a GAL4 protein-upstream activation sequence DNA complex, as shown by (i) coimmunoprecipitation of GAL4 and GAL80 proteins with anti-GAL4 antiserum, (ii) an electrophoretic mobility shift of a GAL4 protein-upstream activation sequence DNA complex upon the addition of GAL80 protein, and (iii) GAL4-dependent binding of GAL80 protein to upstream activation sequence DNA immobilized on Sepharose beads. Anti-GAL4 antisera were raised against a GAL4-URA3 fusion protein, which could be purified to homogeneity in a single step with the use of an affinity chromatographic procedure for the URA3 gene product.

Pubmed ID: 3316976 RIS Download

Mesh terms: DNA, Fungal | DNA-Binding Proteins | Fungal Proteins | Galactose | Gene Expression Regulation | Immunologic Techniques | Macromolecular Substances | Protein Binding | Recombinant Fusion Proteins | Saccharomyces cerevisiae | Transcription Factors

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