FIG. 7.

Deletion analysis of the Orf22 construct. (A) Diagram of N- and C-terminal deletions of Orf22. CEF cells were transfected with 0.3 μg of E2F-Luc reporter plasmid and 0.3 μg of Orf22 construct. Two days after transfection, cells were lysed and assayed for luciferase activity. (B to D) GST-Rb was expressed in bacteria and recovered on glutathione-Sepharose. CEF cells were transfected with each 1.5 μg of Orf22 constructs and GAM-1 plasmid. Two days after transfection, cells were assayed for recombinant protein expression by Western blotting (B and C). Equal amounts of the expressed Myc-tagged protein were incubated with recombinant GST-Rb. Resulting complex formation was analyzed by Western blotting with anti-Myc antibody (D).