FIGURE 4.

Quantitation of Ole1p self-association in vivo membrane with the use of the split luciferase yeast two-hybrid system. A, schematic representation of the plasmid constructs made for expression of Ole1 in the RLUC system in which N- and C-terminal luciferase (Luc) fragments are fused to Ole1p via a 10-amino acid linker (-GGGGSGGGGS-). PC, positive control; NC, negative control; T-T, tail to tail; H-T, head to tail; H-H, head to head; T-H, tail to head. B, assay of the activity of reconstituted RLUC by bimolecular complementation. Relative luminescence units (RLU) are plotted for each construct. Stars designate significant differences from the empty vector (EV), negative control value (*, p < 0.05; **, p < 0.01). Error bars, S.D.