Difference between revisions of "Team:TokyoTech/Description"

Establishment of an artificial co-culture system by communicating between bacteria and human cells

We plan to establish an artificial co-culture system of human cells with bacteria (E. coli) by integration of quorum sensing (cell-to-cell communication between bacteria), His-Asp phosphorelay and cytokinin (a signaling molecule in plant) in this co-existing system as shown in Figure 1

The following give detailed explanations about each step of how the genetic circuit works as shown in Figure 2.

Step1: C8, an autoinducer AHL, is produced and released from E. coli. With increased population of E. coli, the concentration of C8 increases in the culture medium of human cells (C2BBe1) and E. coli.

Step2: C8 binds with TraR expressed in C2BBe1 cells, and p65-TraR-C8 complex promotes the expression of AtIPT4 by the specific signal pathway. The more AtIPT4 expression is promoted, the more Isopentenyladenine (IP) molecules are synthesized in C2BBe1.

Step3: C2BBe1-secreted IP binds with membrane receptor, AHK4, of E. coli.

Step4: IP activates AHK4 and His-Asp phosphorelay delivers phosphorylation signals. Eventually, it leads to activate CPS promoter and downstream mazF is expressed.

Step5: Cleavage of mRNA by a toxin, MazF stops the protein synthesis and the growth of E. coli.