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|1.Sticky Hands]] | [[Chiba/Communication|2.Communication]] | [[Chiba/Quorum_Sensing|3.Size Control]] ) |  [[Chiba/Making Marimo|Making Marimos]] |  [[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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[[Image:Story.PNG|300x300px]][[Image:22s.jpg|150x150px]]<br> (photo taken from Agriculture, Forestry and Fishes Technical Information Society, Japan,  http://www.afftis.or.jp/index.html)<br>
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[[Image:Story.PNG|frame|'''Fig. 4''' Concept|center]]<br>
We aimed to make a spherical gathering of bacteria such like marimo by ordering bacteria go get together and stick to each other.
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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====[[Chiba/Engeneering_Flagella|1.Sticky Tag]]====
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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.<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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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. This image depicts only one tail as flagella (the real bacteria have about ten flagella per cell).]]<br>
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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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====[[Chiba/Communication|2.Communication]]====
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====[[Chiba/Communication|2.Communication Module]]====
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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 sticky hands.<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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違う遺伝子回路を持つ2つの細胞(sender&receiver)に分ける.あらかじめお互い結合しているSenderが,周りのreceiverにシグナルを与えてsticky handsを生やす.そして合体.
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[[Image:Chiba_prj_com.png|frame|left|'''Fig2.''' Bacterial Communication.]]
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[[Image:Chiba_prj_com.png|frame|left|'''Fig. 6''' Bacterial Communication.]]
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====[[Chiba/Quorum_Sensing|3.Size Control]]====
====[[Chiba/Quorum_Sensing|3.Size Control]]====
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Make an '''AHL concentration gradient''' for quorum sensing.  
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Make an '''AHL localized region''' for quorum sensing.  
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[[Image:Chiba_proj_ahlconc.png|frame|left|'''Fig3.''' AHL concentration gradient.]]
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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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===How Our System Works===
===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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[[Image:How our system works chiba.PNG|frame|center|'''Fig. 8''' How Our System Works.]]<br>
#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>
#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>
#Receivers express his/flagella and GFP in high [AHL]; only when they get close to the senders core.<br>
#Receivers express his/flagella and GFP in high [AHL]; only when they get close to the senders core.<br>
#Receivers stick to the senders core and themselves through metal ions.... one after another.<br>
#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.
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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]'''
<!-- SenderやReceiverの色を対応させてみたらどう? by田代 -->
<!-- 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]