Validation of OMV- and OM-enriched proteins.

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Validation of OMV- and OM-enriched proteins. Validation of OMV- and OM-enriched proteins. Candidate ORFs that encode proteins identified in our MS analysis as OMV or OM enriched were cloned into pFD340 with a C-terminal 6×His tag. Constructs were introduced into B. thetaiotaomicron by conjugation, generated strains were grown in TYG medium, and fractions were prepared. Ten micrograms of each fraction was subjected to 12% SDS-PAGE and analyzed by Western blotting using anti-His polyclonal antibodies. (a) OMV-enriched proteins; (b) OM-enriched proteins. The isoelectric point as well as the residues following the lipoprotein attachment cysteine (for lipoproteins) are indicated below the protein name. Ezequiel Valguarnera et al. mSphere 2018; doi:10.1128/mSphere.00559-18