FIG. 7.

Design and expression of MBP fusions of ORF50. (A) Schematic of ORF50/Rta constructs. The numbers refer to amino acids, and the bars underneath ORF50 represent sequences included in the ORF50 mutants identified in the list at the right. The four amino acid sequences at the bottom were inserted at the LR (aa 244 to 275) in the WT, L3P mutant, or chimeric (GCN4 or p53) proteins. ST, serine/threonine-rich sequence; hyd, hydrophobic; DE, acidic amino acid-rich; +++, basic amino acid-rich sequence; *, amino acid position of the mutations in chimeric proteins. (B) MBP fusions of ORF50 stained with Gel Code Blue stain (Pierce). The indicated proteins were expressed and purified from E. coli, as described in Materials and Methods. Equal amounts of protein were displayed by SDS-PAGE, and the gels were fixed and stained with Gel Code Blue (Pierce).