Chiba/Sandbox/Home2

From 2007.igem.org

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:

	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.
	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.
	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.
   • Sequence confirmed
   • Swarm confirmed
   • Flagella strained with anti-flagella antibody
   • Phenotype confirmed
   • Affnity confirmed

more...

Making Communication Module

1. Making Senders
   • AHL > GFP generator (constitutive aiiA) : BBa_T729006
   • AHL > GFP & aiiA generator : BBa_I729005
   • Sensitive AHL > GFP generator : BBa_I729004

[edit] Future Work inverter-aiiA

1. • 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