Tokyo/Works
From 2007.igem.org
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'''Second, we have tried to mathmatically express our model.''' | '''Second, we have tried to mathmatically express our model.''' | ||
- | <br>From simple equations, the equations necessary for our model have been deviated step by step. Though a parameter for production rate of AHL changes cell behavior from conventional mutual-inhibition model, Hill | + | <br>From simple equations, the equations necessary for our model have been deviated step by step. Though a parameter for production rate of AHL changes cell behavior from conventional mutual-inhibition model, Hill coefficients for the promoters are still critical for emergence of bistability.'''([[Tokyo/Works/Formulation |See more.]])''' |
<br>[[Image:Expression3-1.jpg|400px]] | <br>[[Image:Expression3-1.jpg|400px]] | ||
<!--リプレッサーA,B,シグナル分子Sの時間変化を式として表し、安定性解析を行った結果、Hill係数が大切であることがわかった | <!--リプレッサーA,B,シグナル分子Sの時間変化を式として表し、安定性解析を行った結果、Hill係数が大切であることがわかった |
Revision as of 17:16, 26 October 2007
Abstract Concept & Model Requirements Genetic_circuit Works About_our_team
0. Hybrid promoter 1. Formulation 2. Assay1 3. Simulation 4. Assay2 5. Future works
以下Worksのアブストとなる文章に。上下との話のつながりも明記。
How to reach our goal
We have alternatly implemented Wet and Dry experiments to achieve our goal
Our purpose is the construction and development of genetic circuit to establish our "coexistence stability" model. We have alternately employed Dry and Wet approaches. We have drawn a navigational chart on our project by Dry approaches, which has been confirmed and reinforced by the data from Wet approaches.
0. Wet : Hybrid promoter
First, the newly devised promoter sensed the Two inputs is necessary for the Genetic_circuit.
We created a new hybrid promoter and checked its function. (Fig. 1 and 2) This newly devised hybrid promoter regulated by two inputs is essential to the genetic circuit in our project. Since there are no parts available that meet required conditions - activated by AHL and repressed by LacI - among previous publishments as well as BioBrick parts, we started with the design of its sequence. (See more.)
1.Dry : Formulation
Second, we have tried to mathmatically express our model.
From simple equations, the equations necessary for our model have been deviated step by step. Though a parameter for production rate of AHL changes cell behavior from conventional mutual-inhibition model, Hill coefficients for the promoters are still critical for emergence of bistability.(See more.)
2.Wet : Assay1 using "Externally added" materials
Though Hill coefficients are critical for the differentiation, adjustments of Hill coefficients by DNA sequence modification are much more difficult than those of production rate parameters; changes in RBS -35 box, or -10 box allow production efficiency. In order to confirm feasibility of our model, we focused to measure Hill coefficients of our new promoter part.(See more.)
3.Dry : Simulation
Using the data from actual experiments, we simulated behaviors of our model.
Based on stochastic simulation using parameters obtained from our wet experiments, we have confirmed that the whole system becomes unstable when there are only idlers left, and then, they become either of workers and idlers. Also we have determined other parameters necessary for the desired behavior of this system. (See more.)
4.Wet : Assay2 on "Cell-produced" AHL and Expression comparison
Here the simulation results were tested by actual wet experiments.
Whether the parameters obtained in 3 is feasible in actual Wet experiments was tested by focusing on the amount of cell-produced AHL as well as the strength of the promoters. Indication from the result is applied for the next work.(See more.)
5.Future works
Through the experiments mentioned above, we have found that our model can be completed by changing some of the parts employed so far. Continuing such simulation oriented construction of genetic circuit, it will be increasingly necessary to exchange parts. Here we would like to offer useful means for part exchanges.
Based on the results from wet experiments, the next dry approach - analysis and simulation - should be performed, which will in turn be testified and confirmed by wet experiments. Continueing these processes, if time permits, would further sophisticate and complete our model. (See more.)