Chiba/Project Design

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Latest revision as of 05:36, 27 October 2007

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).

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.

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.
Receivers express his/flagella and GFP in high [AHL]; only when they get close to the senders core.
Receivers stick to the senders core and themselves through metal ions.... one after another.
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]

@@ Line 2: / Line 2: @@
 [[Image:chiba_logo.png|center]]
 __NOTOC__
-{| style="border:0;width:100%;" cellpadding="20px" cellspacing="0"
+{| style="border:0;width:100%;font-family:'Trebuchet MS'" cellpadding="20px" cellspacing="0"
 | align="center" |
-[[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]]
+[[Chiba|Home]]<br>
+<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>
+[[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]]
 |}
@@ Line 10: / Line 12: @@
 ==Project Design==
 ===Concept===
-ものがたり<br>
+[[Image:Story.PNG|frame|'''Fig. 4''' Concept|center]]<br>
-[[Image:Story.PNG|300x300px]][[Image:22s.jpg|150x150px]]社団法人　農林水産技術情報協会http://www.afftis.or.jp/index.html<br>
+We aimed to make a spherical gathering of bacteria such like marimo by ordering bacteria go get together and stick to each other.
-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。(We aimed to make a spherical gathering of bacteria such like marimo by scattered bacteria aggregating and sticking each other.)
 ===What our system requires===
 {| style="border:0px;" cellpadding="5px"
 | width="30%" valign="top" |
-====[[Chiba/Engeneering_Flagella|1.Sticky Hand]]====
+====[[Chiba/Engeneering_Flagella|1.Affinity Tag]]====
-Make a '''His-tagged Flagella'''. We aimed to stick bacteria by displaying histidines (which bonds with metal ions) on the flagellar fillament.<br>
+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>
-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>
-[[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>
+[[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>
 | width="30%" valign="top" |
-====[[Chiba/Communication|2.Communication]]====
-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．
+====[[Chiba/Communication|2.Communication Module]]====
-[[Image:Chiba_prj_com.png|frame|left|'''Fig2.''' Bacterial Communication.]]
+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>
+[[Image:Chiba_prj_com.png|frame|left|'''Fig. 6''' Bacterial Communication.]]
 | width="30%" valign="top" |
 ====[[Chiba/Quorum_Sensing|3.Size Control]]====
-Make an '''AHL concentration gradient''' for quorum sensing.
+Make an '''AHL localized region''' for quorum sensing.
-[[Image:Chiba_proj_ahlconc.png|frame|left|'''Fig3.''' AHL concentration gradient.]]
+[[Image:Chiba_proj_ahlconc.png|frame|left|'''Fig. 7''' Controlling AHL diffusing area and the size of Bacteria Marimo.]]
 |}
 ===How Our System Works===
-[[Image:Design.PNG|240x120px]][[Image:Design2.PNG|240x120px]][[Image:Design3.PNG|240x120px]][[Image:Design4.PNG|240x120px]]<br>
+[[Image:How our system works chiba.PNG|frame|center|'''Fig. 8''' How Our System Works.]]<br>
-*実際の形とは違うかもしれないけど、SenderとReceiverの違いをもっとはっきりさせたほうがよいかも？
+#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>
-<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>
+#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>
-#Senderは常にヒスタグをディスプレイした鞭毛を発現させて金属イオンを介してSender同士が接着します。またSenderは常にAHLを合成します。(Senders stick each other with metal ions to express flagella displaying histidines constantly.)<br>
-#AHLが届く範囲にいるReceiverはGFPを発現し、ヒスタグをディスプレイした鞭毛を発現します。<br>
-#ReceiverはSenderの周りに金属イオンを介して接着します。<br>
-#しかしこのままでは大腸菌の集合体が再現なく大きくなってしまいます。そこでReceiverはAHLを分解します。AHLの拡散が抑えられ、鞭毛が生えるReceiverの数が制限されるので大きさが有限に定まります。<br><br>
-#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(Because bacterias of core(A) constantly express what sticks bacterias(X),A stick each other. A constantly send a signal(Y)).<br>
+'''For other strategies to make marimo, visit the official website of [http://chem.tf.chiba-u.jp/igem/ iGEMCHIBA]'''
-#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。(Bacterias receiving Y(B) express GFP and x in range of Y reaching.)<br>
+<!-- SenderやReceiverの色を対応させてみたらどう？　ｂｙ田代 -->
-#BはAの周りに吸着します。(B stick around A.)<br>
-#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。(But bacteria cluster increases in size yet. B degrate Y by expressing what degrate B. Size of Bacteria cluster remain finite to limit number of B receiveing Y by limiting spread of Y.)<br>

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