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Type III secretion system-associated pili found in several plant pathogenic bacteria are required for injection of virulence proteins from bacteria into the plant cells. The possibility to use the type III secretion pilus of Pseudomonas syringae as an epitope display tool was studied. The advantage of the type III secretion pilus, compared with conventional fimbrial epitope display tools, is that the pilin subunits of the type III secretion pilus can auto-assemble into intact pili in vitro. Various peptides were inserted into the type III secretion pilin subunit, and secretion, assembly and surface properties of the modified pili were monitored. It was concluded that the outwards-projecting N-terminal region of the pilin can bear even 43 amino acids insertion. The three-dimensional structure of the epitope, however, can restrict the use of the pilus as an epitope display tool: a β-hairpin structure was poorly tolerated.