Fig. 7. Analysis of the DNA binding properties of the MYB domain of MybE. A C terminus-proximal segment of MybE containing the MYB domain was expressed in E. coli and purified using a C terminal GST tag. Recombinant protein (500 ng) and various competitors (5 pmole) were used for gel retardation assay with a ³²P-labeled 30-mer probe. Extracts were pre-incubated in the presence of: (1) no competitor; (2) the 30-mer wild-type (wt); (3) three mutant forms of the 30-mer (mut 1-3, Fig. 4A and as indicated); (4) duplicated forms of the 30-mer, the dyad region and the C-rich region. The arrow indicates the major complex and we assume that the higher mobility forms, which mirror the behaviour of the major complex, contain degraded forms of the C terminal region of MybE.