Team:Tokyo Tech

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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!-

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. 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-

E. coli was modified to produces ester by addition of 2-MeBuOH or BuOH as a substrate.

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

<flash>file=Tokyotech_ACS.swf|right|quality=best</flash>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!

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…)

3-3. Antibiotic Activation Device

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

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

3-4. mathematical modeling of Artificial Cooperation System

We simulated Artificial Cooperation System and from this result, we predicted our system can work.

(See more…)

4. Full moon inhibits Artificial Cooporation system

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/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