Figure 5

Production of the secretory scFv BL1 following varying gene expression levels. Expression of the gene encoding secretory BL1 was induced with IPTG in Lemo21(DE3) cells grown in the absence and presence of increasing concentrations of rhamnose. BL21(DE3) expressing the gene encoding secretory BL1 and Lemo21(DE3) harboring an empty expression vector were included as controls. A 4 h after induction, cell growth was monitored by measuring A₆₀₀. B Levels and processing of secretory BL1 were monitored using SDS-PAGE followed by immuno-blotting using an α-His antibody. The precursor (p) and the mature (m) form of the protein are indicated. C Using flow cytometry, cell size (forward scatter) and granularity (side scatter) of cells producing secretory BL1 were monitored. D Levels of IbpB, OmpA and MalE were monitored using a combination of SDS-PAGE and immuno-blotting. For OmpA and MalE, the precursor (p) and the mature form (m) of the respective protein are indicated.