Figure 2
The chimeric Intimin-FAST protein is exposed at E. coli cell surface. E. coli MG1655 harbouring pNeae2-FAST (expressing intimin-FAST construct) cells were grown overnight in absence or presence of 0.25 mM IPTG to induce the production of the chimeric Intimin-FAST protein. Bacteria were treated with 0, 5, 10 or 15 µg mL−1 of proteinase K for 15 min at 40 °C before (a) Western blot of whole-cell protein extracts and revelation with a rabbit anti-E-tag primary antibody and a HRP-linked anti-rabbit secondary antibody (The image has been cropped from a larger blot as depicted in Supplementary Information file S8), and (b) immunofluorescence on whole cells using a rabbit anti-E-tag primary antibody and an anti-rabbit Alexa 488 conjugated secondary antibody. All images are displayed with the same constrast. In (a), red stars indicate the position of the full length intimin-FAST. In (b), scale bars = 2 µm.
