Team:Tokyo Tech

From 2010.igem.org

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<h2>0. Project motivation ''-What is humanity?-''</h2><br>
<h2>0. Project motivation ''-What is humanity?-''</h2><br>
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<font size="4" color="#f39700">What do you think humanity is?</font><br>
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<h3>What do you think humanity is?</h3><br>
This is a common question which human have tried for long time. Answer might be different in each person.
This is a common question which human have tried for long time. Answer might be different in each person.
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Then is it really impossible for the other livings to have those?<br>
Then is it really impossible for the other livings to have those?<br>
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<font size="4" color="#f39700">Tokyo Tech 2010 challenged this question. We designed ''E. coli'' with humanity in order to prove that sympathetic doesn’t exist only in human!</font><br>
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<font size="4" color="#cc0033">Tokyo Tech 2010 challenged this question. We designed ''E. coli'' with humanity in order to prove that sympathetic doesn’t exist only in human!</h><br>
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<font size="5" color="#f39700">1. Concept -''E. coli'' with humanity-</font><br>
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<h2>1. Concept -''E. coli'' with humanity-</h2><br>
To make ''E. coli'' with humanity, we decided to engineer ''E. coli'' that behave kindly and have sympathy. Below is a behavior of the ''E. coli'' with humanity we thought.
To make ''E. coli'' with humanity, we decided to engineer ''E. coli'' that behave kindly and have sympathy. Below is a behavior of the ''E. coli'' with humanity we thought.
There are two kinds of cells. In normal conditions, they are competitors for survival. While in critical conditions, such that one is dying, another cell would notice the crisis of dying cell and help her.
There are two kinds of cells. In normal conditions, they are competitors for survival. While in critical conditions, such that one is dying, another cell would notice the crisis of dying cell and help her.
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<font size="5" color="#f39700">2. Apple reporters</font><br>
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<h2>2. Apple reporters</h2><br>
Our ''E. coli'' with humanity gives “apple” as expression of appreciation.
Our ''E. coli'' with humanity gives “apple” as expression of appreciation.
It’s really difficult to make ''E. coli'' producing real apples.However,We succeeded in synthesizing two components of apples.  
It’s really difficult to make ''E. coli'' producing real apples.However,We succeeded in synthesizing two components of apples.  
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<font size="3" color="#f39700">2-1. Colour -Carotenoid synthetic pathway complete!-</font><br>
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<h3>2-1. Colour -Carotenoid synthetic pathway complete!-</h3><br>
[[Image:tokyotech_top1.JPG|250px|right]]This year, our expaned over the project of Cambridge 2009 and completed it.  
[[Image:tokyotech_top1.JPG|250px|right]]This year, our expaned over the project of Cambridge 2009 and completed it.  
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<font size="3" color="#f39700">2-2. Fragrance -We create Apple flavor in ''E. coli''-</font><br>
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<h3>2-2. Fragrance -We create Apple flavor in ''E. coli''-</h3><br>
[[Image:tokyotech_top2.JPG|250px|right]]''E. coli'' was modfied to produces ester by addtion of 2-MeBuOH or BuOH as a substrate.  
[[Image:tokyotech_top2.JPG|250px|right]]''E. coli'' was modfied to produces ester by addtion of 2-MeBuOH or BuOH as a substrate.  
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<font size="5" color="#f39700">3. Artificial Cooporation System</font><br>
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<h2>3. Artificial Cooporation System</h2><br>
In figure1,we explain ''E. coli'' with humanity we want to create. We already explain No.4 in the figure.  
In figure1,we explain ''E. coli'' with humanity we want to create. We already explain No.4 in the figure.  
Next move on to No.1&2&3. In ordr to realize the idea, we constructed a system that named “Artificial Cooperation System"
Next move on to No.1&2&3. In ordr to realize the idea, we constructed a system that named “Artificial Cooperation System"
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<font size="3" color="#f39700">3-1.  Genetic circiut of Artificial Cooperation System</font><br>
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<h3>3-1.  Genetic circiut of Artificial Cooperation System</h3><br>
There are two types of ''E. coli'' in this system. These two types of cells recognize each other by Quorum Sensing.  
There are two types of ''E. coli'' in this system. These two types of cells recognize each other by Quorum Sensing.  
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<font size="3" color="#f39700">3-2. mathematical modeling of  Artificial Cooperation System</font><br>
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<h3>3-2. mathematical modeling of  Artificial Cooperation System</h3><br>
[[Image:tokyotech_top5.JPG|250px|right]]We simulated Artificial Cooporation System and from this result, we predicted our system can work .
[[Image:tokyotech_top5.JPG|250px|right]]We simulated Artificial Cooporation System and from this result, we predicted our system can work .
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<font size="3" color="#f39700">3-3.  Lux repression promoter can work!</font><br>
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<h3>3-3.  Lux repression promoter can work!</h3><br>
[[Image:tokyotech_top3.JPG|250px|right]]In order to construct Artificial Cooporation System, lux activation/repression device plays the key role.<br>
[[Image:tokyotech_top3.JPG|250px|right]]In order to construct Artificial Cooporation System, lux activation/repression device plays the key role.<br>
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<font size="3" color="#f39700">3-4.  Antibiotic Activation Device</font><br>
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<h3>3-4.  Antibiotic Activation Device</h3><br>
[[Image:tokyotech_top4.JPG|250px|right]]We introduced chloramphenicol resistance  gene  the downstream of lux activation promoter. <br>
[[Image:tokyotech_top4.JPG|250px|right]]We introduced chloramphenicol resistance  gene  the downstream of lux activation promoter. <br>
This device activate in the presence of AHL. Sinal molecule activates antibiotic resistance gene.
This device activate in the presence of AHL. Sinal molecule activates antibiotic resistance gene.
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<font size="5" color="#f39700">4. Full moon inhibits Artificial Cooporation system</font><br>
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<h2>4. Full moon inhibits Artificial Cooporation system</h2><br>
[[Image:tokyotech_top6.JPG|250px|right]]In order to have a more intelligible& imaginable system, we tried to link our project to a famous& well-known character ”Wolfman”.<br>
[[Image:tokyotech_top6.JPG|250px|right]]In order to have a more intelligible& imaginable system, we tried to link our project to a famous& well-known character ”Wolfman”.<br>
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<font size="3" color="#f39700">4-1. New series of osmoregulative promoters</font><br>
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<h3>4-1. New series of osmoregulative promoters</h3><br>
[[Image:tokyotech_top7.JPG|250px|left]][[Image:tokyotech_top8.JPG|250px|left]]<br>
[[Image:tokyotech_top7.JPG|250px|left]][[Image:tokyotech_top8.JPG|250px|left]]<br>
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<font size="3" color="#f39700">4-2. LacIMI showed weaker repression than WT</font><br>
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<h3>4-2. LacIMI showed weaker repression than WT</h3><br>
[[Image:tokyotech_top9.JPG|250px|right]]
[[Image:tokyotech_top9.JPG|250px|right]]

Revision as of 16:40, 26 October 2010

iGEM Tokyo Tech 2010 "E.coli with Humanity"

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Contents

0. Project motivation -What is humanity?-


What do you think humanity is?


This is a common question which human have tried for long time. Answer might be different in each person. According to LONGMAN dictionary, humanity means,
“the state of being human rather than an animal or machine. For example, kindness, respect and sympathy towards others.”

Then is it really impossible for the other livings to have those?
Tokyo Tech 2010 challenged this question. We designed E. coli with humanity in order to prove that sympathetic doesn’t exist only in human!</h>


1. Concept -E. coli with humanity-


To make E. coli with humanity, we decided to engineer E. coli that behave kindly and have sympathy. Below is a behavior of the E. coli with humanity we thought. There are two kinds of cells. In normal conditions, they are competitors for survival. While in critical conditions, such that one is dying, another cell would notice the crisis of dying cell and help her. She gives him “apples” as a expression of her appeciation. The relationship like this exists in human beings.


2. Apple reporters


Our E. coli with humanity gives “apple” as expression of appreciation. It’s really difficult to make E. coli producing real apples.However,We succeeded in synthesizing two components of apples.

2-1. Colour -Carotenoid synthetic pathway complete!-


Tokyotech top1.JPG
This year, our expaned over the project of Cambridge 2009 and completed it.

We succeeded in synthesizing zeaxanthin by adding crtZ into crtEBIY likewise astaxanthin by crtZW. We could identify them by TLC. Astaxanthin is the final metabolite of carotenoid synthetic pathway. (See more…)




2-2. Fragrance -We create Apple flavor in E. coli-


Tokyotech top2.JPG
E. coli was modfied to produces ester by addtion of 2-MeBuOH or BuOH as a substrate.

This ester gives off apple fragrance. (See more…)






3. Artificial Cooporation System


In figure1,we explain E. coli with humanity we want to create. We already explain No.4 in the figure. Next move on to No.1&2&3. In ordr to realize the idea, we constructed a system that named “Artificial Cooperation System"




3-1. Genetic circiut of Artificial Cooperation System


There are two types of E. coli in this system. These two types of cells recognize each other by Quorum Sensing. CellA & CellB carry LasI gene and LuxI gene respectively. To see the rescuing process, click on the right figure to start (See more…)




3-2. mathematical modeling of Artificial Cooperation System


Tokyotech top5.JPG
We simulated Artificial Cooporation System and from this result, we predicted our system can work .

(See more…)






3-3. Lux repression promoter can work!


Tokyotech top3.JPG
In order to construct Artificial Cooporation System, lux activation/repression device plays the key role.

Lux repression part has never been measured in iGEM while Lux activation part has measured by many iGEM teams. This year we assayed the activity of lux repression promoter.(See more…)




3-4. Antibiotic Activation Device


Tokyotech top4.JPG
We introduced chloramphenicol resistance gene the downstream of lux activation promoter.

This device activate in the presence of AHL. Sinal molecule activates antibiotic resistance gene. (See more…)






4. Full moon inhibits Artificial Cooporation system


Tokyotech top6.JPG
In order to have a more intelligible& imaginable system, we tried to link our project to a famous& well-known character ”Wolfman”.

In our attempt to produce “wolfcoli”, we introduced light/swiching function to the Artificial Cooperation System. Light sensitive promoter “PompC” and synthetic band detector circuit were used to perform function designed. (See more…)




4-1. New series of osmoregulative promoters


Tokyotech top7.JPG
Tokyotech top8.JPG




We introduced two new series of light sensitive promoters. We also characterized the activity of lacIM1 which is necessary for werewolf System. (See more…)



4-2. LacIMI showed weaker repression than WT


Tokyotech top9.JPG



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