Team:Chiba/System 2

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Contents

Version 2

Overall Circuit

Our second circiut of double click system is called Recognize circuit.
It consists of recognizing system and memorizing system.
Recognizing system can recognize the existence of input.
And Memorizing system can memorize the existence of input in the past for a while.
When the two system work together, bacteria is going to shine.
We hope this circuit works as double click system.

We use AHL for input and regard injection of AHL as clicking. In our project, CLICK controls transcription of overall DNA sequence.
In recognizing input system, there is a cI operator above gfp DNA sequence.cI recognizes the existence of input in this DNA sequence.
In memorizing system, there is an AND Gate with T7ptag and supD. In this case, the AND Gate remembers that there was a click.
There is a hybrid promoter(cI/T7) above gfp DNA sequence. It is regulated by T7 and cI which work as activator and repressor, respectively. On this promoter, repression is stronger than activation. And also, it is low unregulated activation. Working of this promoter depends on recognizing and memorizing system.
detail...


Result

Construction

We've designed DNA sequence like following Figure.

Chiba Sys2.jpg

Based on this sequence, we've also designed vertor.

Chiba ver2.jpg

Prom-luxR&PT7/cI-GFP vector is constructed collaboration with double click system version 1.


Testing

In this system, lux promoter inverter is sensor of input. We've researched Plux inverter and got some results.

Evaluation


Remark&Notes


Conclusion