Chiba/Project Design

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[[Image:chiba_logo.png|center]]
[[Image:chiba_logo.png|center]]
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[[Chiba|Introduction]] | [[Chiba/Project_Design|Project Design]] | [[Chiba/Engeneering_Flagella|Engeneering Flagella]] | [[Chiba/Quorum_Sensing|Quorum Sensing]] | [[Chiba/Goal|Our Goal]] || [[Chiba/Team_Members|Team Members]] | [[Chiba/Members_Only|メンバ連絡簿]]
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[[Chiba|Home]]<br>
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<span style="font-size:120%;font-weight:bold;">[[Chiba/Introduction|Introduction]] | [[Chiba/Project_Design|Project Design]] ( [[Chiba/Engeneering_Flagella|1.Affinity Tag]] | [[Chiba/Communication|2.Communication Module]] | [[Chiba/Quorum_Sensing|3.Size Control]] ) | [[Chiba/Making Marimo|Making Marimos]] |  [[Chiba/Goal|Our Goal]]</span><br>
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[[Chiba/Acknowledgements|Acknowledgements]] | [[Chiba/Team_Members|Team Members]] | [http://chem.tf.chiba-u.jp/igem/ iGEM Chiba Website] | [[Chiba/Members_Only|メンバ連絡簿]]  
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==Project Design==
==Project Design==
===Concept===
===Concept===
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ものがたり<br>
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[[Image:Story.PNG|frame|'''Fig. 4''' Concept|center]]<br>
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[[Image:Story.PNG|300x300px]][[Image:22s.jpg|150x150px]]<br>
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We aimed to make a spherical gathering of bacteria such like marimo by ordering bacteria go get together and stick to each other.
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ばらばらになっている大腸菌を一ヶ所に集めて吸着させて、まりものような球体の大腸菌の集合体を作ることを目指した。
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===How Our System Works===
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[[Image:Design.PNG|240x120px]][[Image:Design2.PNG|240x120px]][[Image:Design3.PNG|240x120px]][[Image:Design4.PNG|240x120px]]<br>
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#核になる大腸菌（A）は常に大腸菌同士が吸着する何か（X）を発現しているのでA同士が吸着します。Aは常に信号（Y）を送ります。<br>
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#Aが送ったYが届く範囲にいるYを受け取る大腸菌（B）はGFPを発現し、Xを発現します。<br>
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#BはAの周りに吸着します。<br>
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#しかしこのままでは大腸菌の集合体が際限なく大きくなってしまいます。そこでBはYを分解する何か（Z）を発現してYを分解します。Yの広がりが抑えられ、Yを受け取るBの数が制限されるので大きさが有限に定まります。<br>
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===What our system requires===
===What our system requires===
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====[[Chiba/Bacteria_Linker|1.Bacteria linker]]====
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====[[Chiba/Engeneering_Flagella|1.Affinity Tag]]====
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Make a '''His-tagged Flagella'''. We aimed to stick bacteria by displaying histidines (which bonds with metal ions) on the flagellar fillament.
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Make a '''His-tagged Flagella'''. We aimed to stick bacteria by displaying histidines (which bonds each other through metal ions) on the flagellar filament.<br>
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鞭毛にヒスタグをディスプレイし，金属イオンを介して，バクテリアを接着することを目指した．<br>
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[[Image:Chiba_stickbacteria.png|frame|left|'''Fig1. ''' His-tagged flagella as a bactria linker. 鞭毛は便宜上１本しか描いてないが実際は1~10本/細胞．]]<br>
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====[[Chiba/Size_Controller|2.Size Controller]]====
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[[Image:Chiba_stickbacteria.png|frame|left|'''Fig. 5 ''' His-tagged flagella as a bactria linker. This image depicts only one tail as flagella (the real bacteria have about ten flagella per cell).]]<br>
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Make an '''AHL concentration gradient''' for quorum sensing.
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[[Image:Chiba_proj_ahlconc.png|frame|left|'''Fig2.''' AHL concentration gradient.]]
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===Genetic Circuit===
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====[[Chiba/Communication|2.Communication Module]]====
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[[Image:Chiba_marimosystem.png|center]]
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Make a '''Bacterial Communication'''. We make 2 types of cell(sender&receiver) having different gene circuit. Senders sticking each other in advance send a signal to receivers and receivers grow affinity tags.<br>
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====Sender====
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[[Image:Chiba_prj_com.png|frame|left|'''Fig. 6''' Bacterial Communication.]]
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[[Image:Send gene circuit.jpg|110x44px]]<br>
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常にFliC-Hisを発現する。常にAHLを合成する。
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====Receiver====
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====[[Chiba/Quorum_Sensing|3.Size Control]]====
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*BBa_T9002(Normal Receiver)
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Make an '''AHL localized region''' for quorum sensing.
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[[Image:BBa T9002.jpg]]<br>
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[[Image:Chiba_proj_ahlconc.png|frame|left|'''Fig. 7''' Controlling AHL diffusing area and the size of Bacteria Marimo.]]
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AHL存在下でGFPを発現する。AHL存在下でFliC-Hisを発現する。
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===How Our System Works===
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[[Image:How our system works chiba.PNG|frame|center|'''Fig. 8''' How Our System Works.]]<br>
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#Senders whose flagella display the histidine tags stick to each other through metal ions. This becomes the core of Bacteria Marimo. The senders also produce AHL to sign the receiver cells.<br>
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#Receivers express his/flagella and GFP in high [AHL]; only when they get close to the senders core.<br>
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#Receivers stick to the senders core and themselves through metal ions.... one after another.<br>
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#As seen, the cluster grows like a snowball. At the same time, the receivers degrade AHL and thus AHL diffusion space around the mixed cluster limited. By controlling the rate of AHL degradation and so on, one can define the size of such Bacteria Marimo.<br>
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*Sensitive Receiver
 
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[[Image:Mutant rec.jpg]]
 
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*Inverter aiiA Receiver
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'''For other strategies to make marimo, visit the official website of [http://chem.tf.chiba-u.jp/igem/ iGEMCHIBA]'''
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[[Image:rec_inv_aiia.jpg|402x45px]]
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<!-- SenderやReceiverの色を対応させてみたらどう？　ｂｙ田代 -->

Latest revision as of 05:36, 27 October 2007

Chiba logo.png

Home
Introduction | Project Design ( 1.Affinity Tag | 2.Communication Module | 3.Size Control ) | Making Marimos | Our Goal
Acknowledgements | Team Members | [http://chem.tf.chiba-u.jp/igem/ iGEM Chiba Website] | メンバ連絡簿


Project Design

Concept

Fig. 4 Concept

We aimed to make a spherical gathering of bacteria such like marimo by ordering bacteria go get together and stick to each other.

What our system requires

1.Affinity Tag

Make a His-tagged Flagella. We aimed to stick bacteria by displaying histidines (which bonds each other through metal ions) on the flagellar filament.

Fig. 5 His-tagged flagella as a bactria linker. This image depicts only one tail as flagella (the real bacteria have about ten flagella per cell).

2.Communication Module

Make a Bacterial Communication. We make 2 types of cell(sender&receiver) having different gene circuit. Senders sticking each other in advance send a signal to receivers and receivers grow affinity tags.

Fig. 6 Bacterial Communication.

3.Size Control

Make an AHL localized region for quorum sensing.

Fig. 7 Controlling AHL diffusing area and the size of Bacteria Marimo.

How Our System Works

Fig. 8 How Our System Works.

  1. Senders whose flagella display the histidine tags stick to each other through metal ions. This becomes the core of Bacteria Marimo. The senders also produce AHL to sign the receiver cells.
  2. Receivers express his/flagella and GFP in high [AHL]; only when they get close to the senders core.
  3. Receivers stick to the senders core and themselves through metal ions.... one after another.
  4. As seen, the cluster grows like a snowball. At the same time, the receivers degrade AHL and thus AHL diffusion space around the mixed cluster limited. By controlling the rate of AHL degradation and so on, one can define the size of such Bacteria Marimo.


For other strategies to make marimo, visit the official website of [http://chem.tf.chiba-u.jp/igem/ iGEMCHIBA]

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