Figure 7.

Physical Interaction between AhSLF-S₂ and PhS₃-RNase.

(A) Yeast cells containing various combinations of BD and AD fusions were tested for their growth on -Leu/-Trp/-His/-Ade dropout media. Plasmid pGBKT₇ with various AD:constructs and plasmid pGADT₇ with various BD:constructs were used as negative controls.

(B) The strains were grown further to test for expression of the β-galactosidase reporter gene.

(C) A pull-down assay for the physical interaction between AhSLF-S₂ and PhS₃-RNase. Ni-NTA resin and the purified fusion proteins of His-AhSLF-S₂-C were incubated with the style extract of the S₃S₃ line of P. hybrida (Ph Style Extracts). Bound proteins were pulled down with Ni-NTA resin, eluted with the lysis buffer, separated by 12% SDS-PAGE, transferred to membranes, and analyzed by immunoblotting using the anti-S-RNase antibody (top panel). Style extracts from Antirrhinum (S₂S₅) (Ant Style Extracts) were also included as a control. Input style total protein was used as a positive control (bottom panel). Molecular mass markers are indicated in kilodaltons.