Part:BBa_K750009

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

We had took a pre-experiment before we did the fluorescence test. We useed TD1.0 as the experimental subject, and inducted it by 0.1mM, 1.0mM, 10mM arabinose. From this figure, we ensured that 0.1mM arabinose induct the circuit best. So we finally chose 0.1mM arabinose for fluorescence test later. Then we did the fluorescence test of TD1.0 Prepared with its control, fluorescence value of TD1.0 arrived at a much higher degree. That means the GFP has been produced well, the whole circuit are working well. At the same time, we did fuorescence test of another two circuit we had finished construction:TD0.6(BBa_K750010), TD0.01(BBa_K750012). In order to prove that there is a time delay phenomenon when these 3 circuit working, we put curves of them into one figure: We chose the value of fluorescence 5000 to compare. Here is a column chart to display how long these 3 circuit take to arrive the value 5000: And we can find that there surely a time delay phenomenon when these 3 circuit working together. They are working as we designed.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 125
  Illegal NheI site found at 1069
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 793
  Illegal BamHI site found at 65
  Illegal BamHI site found at 1009
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 2024
  Illegal BsaI.rc site found at 2751