Figure 8.

Interaction between OSH15 and Rice KNOX Proteins.

(A) Quantitative yeast two-hybrid assay with OSH15 and rice KNOX (OSH) proteins. For the bait construct, OSH15 was fused to the GAL4 DNA binding domain (GAL4-DB). For the prey constructs, OSH proteins were fused to the GAL4 activation domain (GAL4-AD). The pairwise combination of the bait and prey constructs for each row is shown at left. Relative β-galactosidase (LacZ) activity was calculated by standardizing each LacZ activity with that of intact OSH15/OSH15. We set the activity of OSH15/OSH15 at 100. Error bars represent the standard deviations calculated from three independent yeast clones. The combinations of OSH15–empty vector and OSH15–T antigen are negative controls of interaction, whereas the combination of p53–T antigen is a positive control.

(B) In vitro pull-down assay with ³⁵S-methionine–labeled OSH15. The fusion proteins consisted of OSH proteins and GST: GST-OSH1 (lane 3); GST-OSH15 (lane 4); GST-OSH43 (lane 5); GST-OSH6 (lane 6); and GST-OSH71 (lane 7). The input protein, ³⁵S-methionine–labeled OSH15, was loaded onto the gel alone as a control (lane 1). GST alone was used as negative control for interaction (lane 2).