Team:Tokyo Tech/Project
From 2008.igem.org
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=== Part 3 === | === Part 3 === | ||
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Revision as of 10:59, 21 October 2008
Home | The Team | The Project | Parts Submitted to the Registry | Modeling | Notebook |
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(Or you can choose different headings. But you must have a team page, a project page, and a notebook page.)
Our project
We want to make a Coli Touch!!
→We try to construct a bacterial ‘touch panel’ which is colored by the pressure.
We name it Coli touch.
Why pressure?
Example of input style to genetic circuit
- Heat: thermal gradient
- Small molecule: concentration gradient
- Light: strength depend on location of container
Difficulty in uniform introduction
Press! Confirmatory experiment
Prospect of technological application!!
Pressure
inducible promoter - Plac
The affinity of LacI for the lac operator is known to decrease due to a tetramer to dimer transition of LacI
![[サンプルの絵]](https://static.igem.org/mediawiki/2008/b/bd/Test.jpg)
Strategy
![[サンプルの絵]](https://static.igem.org/mediawiki/2008/c/cf/Test2.jpg)
Equipment. Instrument for pressure experiment
![](https://static.igem.org/mediawiki/2008/a/a3/Tech_kaatu.jpg)
Fig 4: Fig4-1 is a pressure vessel. Fig4-4 is a pressurizing system. Fig4-2 show a inside of the vessel. The inside is filled with water and the withstand pressure container is put into the inside. Fig4-3 show a water bath placed some vessel.
Strategy
1.Construction
![](https://static.igem.org/mediawiki/2008/6/63/Tech_constraction.jpg)
Confirmatory
experiment
![](https://static.igem.org/mediawiki/2008/6/6b/Tech_plac_g1.jpg)
Fig1:Pressure-response of Plac-GFP under several pressures measured by flow cytometer
![](https://static.igem.org/mediawiki/2008/e/e5/Tech_pic_30.jpg)
![](https://static.igem.org/mediawiki/2008/a/a2/Tech_pic_rp.jpg)
![](https://static.igem.org/mediawiki/2008/f/f6/Tech_pic_nega.jpg)
Fig2-1:Plac-GFP under 30MPa Fig2-2:Plac-GFP under 0.1MPa Fig2-3:⊿-GFP under 30MPa
Stratagy 2. Basic device
![](https://static.igem.org/mediawiki/2008/9/9d/Tech_youki.jpg)
Stratagy 3. Development of low pressure inducible
promoter
We performed random mutagenesis to lac promoter and tried to screen promoters with FACS.
Strategy for Isolation of promoters
Future
work
• try Step 2 and 3
• characterize isolated promoters
4.Genetic
toggle switch to implement rewritable function
・ genetic toggle switch
- Writable function
Plac is induced under 30 MPa pressure
(PL is not induced by 30 MPa pressure)
Express GFP, CI, and CI represses PL
→ bright !!
- Erasable function
PL is induced by heat
Express LacI, Plac represses GFP
→ go out little by little…
Future work
- Identify pressure model parameter from experimental data
- Simulate pressure induction by pressure model and estimate
promoter-RBS strength to implement rewritable function
Mathematical model
Contents |