The DNA-binding domain of c-Myb: overexpression and NMR characterization

Abstract

The DNA-binding domain of the Drosophila melanogaster c-Myb protein, 160 residues long, containing three conserved imperfect repeats of nearly 50 residues each has been overexpressed in E. coli strain BL21(DE3). The protein is expressed to the extent of atleast 20% of the total cellular protein. It has been purified by a four-step protocol developed in the laboratory. The protein has been characterized by various 2D NMR experiments. Several specific amino acids have been identified. A three-dimensional NMR spectrum has been recorded to achieve dispersion of cross-peaks into different 2D planes.