Chiba/Sandbox/Home2

From 2007.igem.org

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(Making Affinity Tags)
(Making Affinity Tags)
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==Achievements Overview==
==Achievements Overview==
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===Making Affinity Tags===
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===[[Chiba/Engeneering_Flagella|Making Affinity Tags]]===
[[Image:Chiba_flichisgene.png|frame|'''Fig.''' The short peptide with six histidine (“His-Tag”) was inserted into the fliC D3 domain.]]
[[Image:Chiba_flichisgene.png|frame|'''Fig.''' The short peptide with six histidine (“His-Tag”) was inserted into the fliC D3 domain.]]
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[[Image:Beads-Adsorption_Colony-Number.jpg|frame|'''Fig.'''Beads adsorption assay.]]
 
#Make the affinity tag by inserting the his-tag into the flagellar fillament.
#Make the affinity tag by inserting the his-tag into the flagellar fillament.
#*[[Image:Chiba_check.png]] Sequence confirmed
#*[[Image:Chiba_check.png]] Sequence confirmed
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#*[[Image:Chiba_check.png]] Flagella strained with anti-flagella antibody
#*[[Image:Chiba_check.png]] Flagella strained with anti-flagella antibody
#*[[Image:Chiba_check.png]] Phenotype confirmed
#*[[Image:Chiba_check.png]] Phenotype confirmed
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#*[[Image:Chiba_check.png]] Affnity confirmed ([[Chiba/Engeneering_Flagella|more...]]])
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#*[[Image:Chiba_check.png]] Affnity confirmed
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[[Chiba/Engeneering_Flagella|more...]]
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Revision as of 03:43, 27 October 2007

Chiba logo.png

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

Chiba's Project

Our iGEM project is to make a Marimo-ish gathering of bacteria. Marimo is a green spherical shaped algae (shown in Fig.1), which is a popular living organism in Japan as a National Treasure.(<because of its shape?) Motivation.
(more...)

Design

Our marimo bacteria requres as follows:

Chiba stickbacteria.png

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.

Chiba prj com.png

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.

Chiba proj ahlconc.png

3.Size Controler. Make an AHL localized region for quorum sensing.

これを組み合わせて以下のように毬藻をつくる:

Achievements Overview

Making Affinity Tags

Fig. The short peptide with six histidine (“His-Tag”) was inserted into the fliC D3 domain.
  1. Make the affinity tag by inserting the his-tag into the flagellar fillament.
    • Chiba check.png Sequence confirmed
    • Chiba check.png Swarm confirmed
    • Chiba check.png Flagella strained with anti-flagella antibody
    • Chiba check.png Phenotype confirmed
    • Chiba check.png Affnity confirmed

more...

Making Communication Module

  1. Making Senders
    • 1.---
    • 2.---
  2. Making Receivers
    • 1.---
    • 2.---

Controlling Size

  1. Improve Sender
    • 前駆体合成酵素metKを入れてみた.>結果?
  2. Improve Receiver
    • LuxRの感度が良くなるmutation(論文より)を入れてみた.>結果
  3. Localize AHL
    • aiiAその1の回路(〜〜になるはず)>結果
    • aiiAその2の回路
    • その3

Making Marimo

Remaining Task(Work)

Our Goal

Future