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. 2006 Aug;188(16):5865–5877. doi: 10.1128/JB.00060-06

FIG. 2.

FIG. 2.

Expression of FLAG-tagged HP0546 in three different bacterial hosts, E. coli, A. tumefaciens, and H. pylori, reveals that the protein is localized in the insoluble bacterial fractions and is processed. (A) Expression of HP0546 (of strain 26695) in E. coli MC1061. (B) Expression of HP0546 in A. tumefaciens NTREB1. (C) Expression of HP0546 in H. pylori 26695. Proteins (5 μg per lane) were separated on 16.5% Tris-Tricine SDS-PAGE gels, and Western blots were developed with anti-FLAG tag M2 monoclonal antibody. Only proteins from insoluble fractions after ultrasonication are shown, since no signals were detected in the soluble fractions. Lanes 1, insoluble fractions, which show HP0546 protein expression (plasmid constructs used for HP0546 expression are pCJ206 [A], pCJ207 [B], and pCJ205 [C]); lanes 2, insoluble fractions of negative controls without expression of specific FLAG-tagged proteins (plasmids used as negative controls are pCJ201-0 [A], pCJ203-1w [B], and pCJ201-0 [C]). Molecular masses of standard proteins are indicated on the left; arrows indicate proteins specifically detected with the anti-FLAG tag antibody.