Figure 2.
Constitutive, stable, and nontoxic expression of synthetic adhesins from the chromosome of E. coli. (a) Scheme of a SA gene fusion integrated in the flu gene of E. coli chromosome under the control of the constitutive promoter PN25. (b) Flow cytometry analysis of E. coli EcM1luxΔflu, EcM1luxSAgfp, or EcM1luxSAtir bacteria. Histograms show the fluorescence intensity of bacteria stained with anti-myc mAb and secondary anti-mouse IgG-Alexa 488. (c) Growth curves of bacterial cultures of the same strains shown in panel b. (d) Western blot analysis of the expression of SAs in EcM1luxSAgfp and EcM1luxSAtir strains grown in LB cultures for the indicated time. SAs were immunodetected with anti-myc tag mAb. Cytoplasmic GroEL chaperone was used as loading control (lower panel) and was detected with anti-GroEL mAb.