Chiba/Project Design

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[[Image:chiba_logo.png|center]]
[[Image:chiba_logo.png|center]]
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{| style="border:0;width:100%;font-family:'Trebuchet MS'" cellpadding="20px" cellspacing="0"
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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|150x150px]]⇒[[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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大腸菌を一ヶ所に集め、まりものような球体を作ることを考えた。
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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'''. 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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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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[[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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Make an '''AHL concentration gradient''' for quorum sensing.
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（絵）
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===Genetic Circuit===
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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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[[Image:Chiba_marimosystem.png|center]]
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====Sender====
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====[[Chiba/Communication|2.Communication Module]]====
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[[Image:Send gene circuit.jpg|110x44px]]<br>
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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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常にFliC-Hisを発現する。常にAHLを生産する。
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====Receiver====
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[[Image:Chiba_prj_com.png|frame|left|'''Fig. 6''' Bacterial Communication.]]
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*BBa_T9002(Normal Receiver)
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[[Image:BBa T9002.jpg]]<br>
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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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*Sensitive Receiver
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====[[Chiba/Quorum_Sensing|3.Size Control]]====
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[[Image:Mutant rec.jpg]]
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Make an '''AHL localized region''' for quorum sensing.
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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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*Inverter aiiA Receiver
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[[Image:rec_inv_aiia.jpg|402x45px]]
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===How Our System Works===
===How Our System Works===
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[[Image:Design.PNG|400x200px]]<br>
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[[Image:How our system works chiba.PNG|frame|center|'''Fig. 8''' How Our System Works.]]<br>
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①Senderは常にヒスタグ付きの鞭毛が生えているのでニッケルイオンを仲介して大腸菌同士が吸着します。Senderは常にAHLを生産します。<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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②Senderが生産したAHLが届く範囲にいるReceiverはGFPを発現し、ヒスタグ付きの鞭毛を生やします。<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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③ヒスタグ付きの鞭毛が生えたReceiverはニッケルイオンを仲介としてSenderの周りに吸着します。<br>
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#Receivers stick to the senders core and themselves through metal ions.... one after another.<br>
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④ReceiverはaiiAを発現してAHLを分解します。AHLが届く範囲が狭まり、鞭毛が生えるReceiverの数が制限され大腸菌の集合体の大きさが制限されます。<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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'''For other strategies to make marimo, visit the official website of [http://chem.tf.chiba-u.jp/igem/ iGEMCHIBA]'''
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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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