FIG 7.

Determination of the influence of gp79 on the expression of ORF34₅₂ using the inducible tnaN promoter. (A and B) The cured strain N. magadii L13 bearing pRo-5-ptna-bgaH was grown in minimal medium and either induced by tryptophan addition every 24 h (open circles) or uninduced (filled squares). Samples were taken at the indicated time points, specific BgaH activity was measured (A), and the optical density at 700 nm was determined (B). (C and D) The cured strain N. magadii L13 bearing pNB102-ORF34₅₂ and pRo-5-ptna-ORF79 was grown in minimal medium and either induced by tryptophan addition every 24 h (open circles; cont.) or uninduced (filled squares; un.) or induced only once 72 h after inoculation (filled circles; once). Samples were taken at the indicated time points, the optical density of the culture was measured at 700 nm (C), and total protein extracts were prepared. Protein samples were used for Western blot analysis using polyclonal antibodies against gp34 (D). (E) The cured strain N. magadii L13 bearing pNB102-Mtase and pRo-5-ptna-ORF79 was grown in minimal medium and either induced by tryptophan addition every 24 h (open circles; cont.) or uninduced (filled squares; un.). Total protein samples were taken at the indicated time points and used for Western blot analysis using a polyclonal antibody against M.NmaϕCh1-I.