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Residues that are required for EspP processing are also essential for the processing of B. pertussis autotransporters. BL21-CodonPlus(DE3)−RIL was transformed with a plasmid encoding HA-tagged Prn or the indicated mutant (A), or His-tagged BrkA or the indicated mutant (B) and grown in LB. Autotransporter synthesis was induced by incubating cultures with 100 μM IPTG for 2 h, and passenger domain cleavage was assayed by Western blot using anti-HA or anti-His tag antisera.
