Team:Paris Bettencourt/Xylose diffusion

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Team IGEM Paris 2011


The aim of this step is to diffuse a small metabolite through the nanotubes. The metabolite choosen here is the xylose system, because it has a strong enhancement activity via the promoter.

First, we have to hijack the hierarchy of sugar digestion, since we are working on a LB with glucose, and because we will add IPTG in the medium. To make the system independant, we have choosen to knock out the CRP gene, to short-cut the regulation of glucose over xylose, and ... to shortcut the lactose regulation.

To prevent the degradation of the xylose, we also have to KO the XylAB genes, but keep the importation system XylFGH working. The emitor cell is incubated a few hours in presence of both glucose and xylose, so that the xylose is accumated within the cell at high concentration. Then the emittor cells are mixed with the receptor cells. When the bridge is established between the cells, the xylose diffuse through, and activate the responce of the GFP.

The general scheme is given below: