To verify our construct was being produced by the cells we used fluorescence microscopy to detect the mCherry protein tagged to our construct. As we can see in the figure below, after 10 ms of red light exposure, the induced cell clearly emits a lot of light whereas the uninduced cell barely emits any light (the slight light emission they produce is due to promoter leakage). In essence, our construct is emitted upon induction.

To verify our construct was being secreted into the media we performed a western blot in which we detected our construct with an anti-polyhistidine antibody. As we can see in the figure below, when induced, our construct is being produced by the cells (induced pellet) and is being secreted into media (induced 40 uL supernatant). The presence of the construct in the uninduced cells could be due to promoter leakage. In essence, our construct is successfully secreted into the media.

Between the wiki freeze and the Jamboree in Boston, we plan on finishing the cloning of the antibody scFv in our purification construct, and provide evidence for antibody scFv function by binding it to the Salmonella protein DADH. Be prepared for exciting results!