Fig. 5.

Identification of proteins that interact with PilG and PilH in Xcc. a Western blotting of the eluted 3 × Flag::PilG fusion protein and the eluted 3 × Flag::PilH fusion protein. After Co-immunoprecipitation, a western blot assay was performed for the eluted 3 × Flag::PilG (or 3 × Flag::PilH)fusion protein and the control. b Candidate target proteins of PilG and PilH from two MS-coupled co-IP experiments. c The bacterial two-hybrid experiment showed that PilG and PilH interacted with candidate target proteins. The reporter strain XL1-Blue MRF′ with different plasmid pairs was grown on no selective plates and double-selective indicator plates containing 5 mM 3-AT and 12.5 μg ml^− 1 streptomycin. Protein-protein interactions activated the expression of addA and HIS3 genes within the reporter gene cassette of the reporter strain, resulting in resistance to streptomycin and 3-AT