Difference between revisions of "Part:BBa K750010"

Revision as of 18:40, 25 September 2012

TIME DELAY 0.6:LuxI(RBS0.6)->LuxR->LuxPR->GFP

Description

This part contains BBa_K750002 and BBa_K750007. Arabinose will activate the promoter Pbad, which will cause production of protein LuxI and LuxR. The protein LuxI will synthesize a small, diffusible signaling molecule acyl-homoserinelactone(AHL). The AHL accumulates as the cell density increases. At sufficiently high concentrations, it binds the LuxR, which induces the expression of our reporter GFP under the control of a promoter lux pR. Expression of LuxI will be affected when strength of RBS before LuxI gene is changed. When expression of LuxI is weak, speed of AHL production will go down, that means it costs longer time for AHL-LuxR to arrive the activation thrshold of the promoter LuxPR. In the time delay part of our project E.Lumoli, we designed 4 circuit with RBSes of different strength in the LuxI producer to construct a time delay device. Other time delay parts: TD1.0(BBa_K750009), TD0.3(BBa_K750011), TD0.01(BBa_K750012).

Performance

TIME DELAY1.0:LuxI(RBS1.0)->LuxR->LuxPR->GFP

Description

This part contains BBa_K750001 and BBa_K750007. Arabinose will activate the promoter Pbad, which will cause production of protein LuxI and LuxR. The protein LuxI will synthesize a small, diffusible signaling molecule acyl-homoserinelactone(AHL). The AHL accumulates as the cell density increases. At sufficiently high concentrations, it binds the LuxR, which induces the expression of our reporter GFP under the control of a promoter lux pR. Expression of LuxI will be affected when strength of RBS before LuxI gene is changed. When expression of LuxI is weak, speed of AHL production will go down, that means it costs longer time for AHL-LuxR to arrive the activation thrshold of the promoter LuxPR. In the time delay part of our project E.Lumoli, we designed 4 circuit with RBSes of different strength in the LuxI producer to construct a time delay device. And this part, TD1.0, reacts faster than other circuits we designed(TD0.6:BBa_K750010,TD0.01:BBa_K750012).

Performance

Figure 1. The optimization of the arabinose induced concentration added into the TD1.0 including 0, 0.1, 1.0, 10 mM arabinose.

Figure 2. Fluorescence curves of TD0.6 induced by 0.1mM arabinose and TD1.0 without arabinose.

Figure 3. Fluorescence curves of TD0.01 induced by 0.1mM arabinose and TD0.01 without arabinose.

Figure 4. Fluorescence curves of TD1.0 induced by 0.1mM arabinose and TD0.6 without arabinose

Figure 5. Fluorescence curves of the induced TD0.01, TD0.6, TD1.0 and the control BL21

Figure 6. Experiemently measured the average time of the three cultures’ fluorescence up to 5000 after induction.

Sequence and Features