Article Title: The cysteine-rich exosporium morphogenetic protein, CdeC, exhibits self-assembly properties that lead to organized inclusion bodies in Escherichia coli
Figure Lengend Snippet: Effect of recombinant strain type in the soluble expression and structural organization of CdeC. A) Recombinant CdeC expression in E. coli BL21 (DE3) pRIL and SHuffle T7 strains carrying pARR19 were induced with 0.5 mM IPTG for 16 h at 37°C. Cells were collected and lysed in soluble (S) and insoluble (I) lysis buffer, electrophoresed, and analyzed by Western blot as described in the Methods section. Each lane was loaded with 2 μg of protein lysate. Molecular mass (kDa) markers are indicated at the left side of the panels, and molecular mass of the detected His-tagged immunoreactive bands are indicated at the right side of the panels. B) Thin sections of E. coli BL21 (DE3) pRIL and E. coli SHuffle T7 expressing CdeC from C. difficile R20291 were analyzed by transmission electron microscopy as described in the Method section. Representative micrographs of several E. coli cells are shown in the upper panel. The middle panel shows selected individual cells, and the lower panel displays a magnified view of the thin section of inclusion bodies inside E. coli BL21 (DE3) pRIL or SHuffle T7. When corresponding, lamellae-like formation in CdeC is indicated with blue arrowheads.
Article Snippet: Also, plasmid pARR19 was transformed into E. coli strains BL21(DE3) pRIL and SHuffle T7 ( ).
Techniques: Recombinant, Expressing, Lysis, Western Blot, Transmission Assay, Electron Microscopy