FIG. 5.

Colony filter lift assay to identify the specificity of interaction between FliA and FlgM. S. cerevisiae Y187 carrying recombinant plasmids pAS2-fliA and pACT-flgM (sector A) and parental plasmids pAS2-1 and pACT2 (sector B) were grown on yeast synthetic dropout medium (−Trp/−Leu) and transferred to a Whatman no. 5 paper filter. Cells were permeabilized by freeze-thaw treatment of the filters. The filters carrying the cells were then placed over filters presoaked with Z-buffer-X-Gal (5-bromo-4-chloro-3-indolyl-β-d-galactopyranoside) solution. A blue color revealed within 30 min was considered positive. Assays were repeated three times with at least four independent transformants.