Tokyo/Genetic circuit

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Revision as of 19:35, 25 October 2007

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Genetic Circuit

Fig.1 is an outline of the genetic circuit in our model, designed to realize our Concept & Model.

All cells, wokers and idlers, have THE SAME genetic circuit (just as 'society' members have practically the same genetic backgrounds).

Fig.1: Genetic Circuit
All cells, wokers and idlers, have the same genetic circuit. Workers expresses RFP; while, idlers GFP.

Workers (State-A)

As shown in Fig. 2, an A-state cell, in which the promoter Pa is "on," expresses the repressor Ra, luxI, and RFP. The repressor Ra "turns off" the promoter Pb and prevents from changing the cell states. This cell also expresses luxI, which makes AHL, cell-cell communication molecules. Downstream of these genes, there is an RFP gene; therefore, we would be able to detect state-A cell using flow cytometer.

Fig.2: Workers (State-A)
A worker, an A-state cell expresses the Pa’s downstream genes including the repressor Ra, luxI, and RFP. The Pb and its downstream genes for idlers are positively repressed.

Idlers (State-B)

Fig. 3 shows a B-state cell, in which promoter PB is "turned on" in the presence of AHL, and repressor Rb and GFP are expressed. Repressor Rb represses the expression of promoter Pa and thereby RFP; therefore, fluorescence of GFP is used to detect state-B cells.

If Workers are removed...

As shown in Fig 4, if A-state cells are removed, B-state cells sense the absense of A-cells. they respond to the decrease in AHL, stop producing repressor Rb. Now that repressions of Pa by Rb as well as that of Pb by Ra are declined, either states cannot be clearly expressed, as if the cells were reset.

Fig.3: Idlers (State-B)
In the presence of enough AHL, an idler, a B-state cell expresses the Pb’s downstream genes including the repressor Rb and GFP. The Pa and its downstream genes for workers are positively repressed.

Fig.4: If Workers are removed...
If workers are removed, AHL concentration decreases, which decreases activation of promoter Pb. With less gene expression downstream of Pb, the B-state becomes unstable.

Hybrid promoter

To realize such a genetic circuit, Pb should be "Hybrid promoter" regulated by LuxR and LacI as shown in the figures.