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Mutation of residues 1066 and 1081 reduces the stability of the EspP β domain. AD202 transformed with pRLS5 (Ptrc-espP) or a pRLS5 derivative encoding the indicated EspP mutant were harvested after the addition of IPTG. Cell membranes were isolated and heated at the indicated temperature before proteins were resolved by SDS/PAGE. The cleaved EspP β domain was detected by Western blot using an antiserum generated against a C-terminal peptide.
