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Team:Tokyo Tech
From 2010.igem.org
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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> | ||
<font size="4" color="#1648e0">Tokyo Tech 2010 challenged this question.<br>We designed ''E. coli'' with humanity in order to prove that sympathetic doesn’t exist only in human!</font><br> | <font size="4" color="#1648e0">Tokyo Tech 2010 challenged this question.<br>We designed ''E. coli'' with humanity in order to prove that sympathetic doesn’t exist only in human!</font><br> | ||
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<h2>1. Concept -''E. coli'' with humanity-</h2><br> | <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.<br> |
| + | 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. | ||
She gives him “apples” as a expression of her appeciation. | She gives him “apples” as a expression of her appeciation. | ||
The relationship like this exists in human beings. | The relationship like this exists in human beings. | ||
| - | [[Image:Tokyotech_top10.jpg|center|640px]] | + | [[Image:Tokyotech_top10.jpg|center|640px|thumb|fig.1-1 1.They are competitors. 2.One is dying, another cell notices it. 3.fine cell rescues dying cell. 4.Dying cell recovers snd she gives "Apples" as expression of appreciation]] |
<h2>2. Apple reporters</h2><br> | <h2>2. Apple reporters</h2><br> | ||
| - | Our ''E. coli'' with humanity gives | + | Our ''E. coli'' with humanity gives “apples” 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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<h3>2-1. Colour -Carotenoid synthetic pathway complete!-</h3><br> | <h3>2-1. Colour -Carotenoid synthetic pathway complete!-</h3><br> | ||
| - | [[Image:tokyotech_top1.JPG|250px|right]]This year, our | + | [[Image:tokyotech_top1.JPG|250px|right]]This year, our expanded 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. | 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. | Astaxanthin is the final metabolite of carotenoid synthetic pathway. | ||
| - | (See more…) | + | [Team:Tokyo_Tech/Project/Apple_Reporter (See more…)] |
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<h3>2-2. Fragrance -We create Apple flavor in ''E. coli''-</h3><br> | <h3>2-2. Fragrance -We create Apple flavor in ''E. coli''-</h3><br> | ||
| - | [[Image:tokyotech_top2.JPG|250px|right]]''E. coli'' was | + | [[Image:tokyotech_top2.JPG|250px|right]]''E. coli'' was modified to produces ester by addition of 2-MeBuOH or BuOH as a substrate. |
This ester gives off apple fragrance. | This ester gives off apple fragrance. | ||
| - | (See more…) | + | [Team:Tokyo_Tech/Project/Apple_Reporter2 (See more…)] |
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<h2>3. Artificial Cooporation System</h2><br> | <h2>3. Artificial Cooporation System</h2><br> | ||
| - | + | At section 1, we explained ''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 | + | Next move on to No.1&2&3. In order to realize the idea, we constructed a system that named “Artificial Cooperation System" |
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<h3>3-1. Genetic circiut of Artificial Cooperation System</h3><br> | <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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| - | <h3>3- | + | <h3>3-2. 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> | ||
Lux repression part has never been measured in iGEM while Lux activation part has measured by many iGEM teams. | Lux repression part has never been measured in iGEM while Lux activation part has measured by many iGEM teams. | ||
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| - | <h3>3- | + | <h3>3-3. 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. | + | This device activate in the presence of AHL. Signal molecule activates antibiotic resistance gene. |
(See more…) | (See more…) | ||
| - | + | <h3>3-4. mathematical modeling of Artificial Cooperation System</h3><br> | |
| + | [[Image:tokyotech_top5.JPG|250px|right]]We simulated Artificial Cooperation System and from this result, we predicted our system can work. | ||
| + | (See more…) | ||
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<h2>4. Full moon inhibits Artificial Cooporation system</h2><br> | <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> | ||
| - | In our attempt to produce “wolfcoli”, we introduced light/ | + | In our attempt to produce “wolfcoli”, we introduced light/switching function to the Artificial Cooperation System. Light sensitive promoter “PompC” and synthetic band detector circuit were used to perform function designed. |
(See more…) | (See more…) | ||
Revision as of 00:51, 27 October 2010
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!
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 “apples” 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!-
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. [Team:Tokyo_Tech/Project/Apple_Reporter (See more…)]
2-2. Fragrance -We create Apple flavor in E. coli-
This ester gives off apple fragrance. [Team:Tokyo_Tech/Project/Apple_Reporter2 (See more…)]
3. Artificial Cooporation System
At section 1, we explained 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 order 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. Lux repression promoter can work!
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…)
3-3. Antibiotic Activation Device
This device activate in the presence of AHL. Signal molecule activates antibiotic resistance gene. (See more…)
3-4. mathematical modeling of Artificial Cooperation System
(See more…)
4. Full moon inhibits Artificial Cooporation system
In our attempt to produce “wolfcoli”, we introduced light/switching 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
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
