Chiba/Sandbox/Home2
From 2007.igem.org
(Difference between revisions)
| Line 61: | Line 61: | ||
===[[Chiba/Quorum_Sensing|Controlling Size]]=== | ===[[Chiba/Quorum_Sensing|Controlling Size]]=== | ||
#Improve Sender | #Improve Sender | ||
| - | #* | + | #*[[Image:Chiba_check.png]] Overexpressed the metK (the AHL precursur synthesis enzyme) in the hope to increase AHL synthesis. (Turned out not to work) |
#Improve Receiver | #Improve Receiver | ||
| - | #* | + | #*[[Image:Chiba_check.png]] Inserted 2 mutations in LuxR which is known from the paper to increase activity. |
#Localize AHL | #Localize AHL | ||
| - | #* | + | #*[[Image:Chiba_check.png]] Tested the GFP expression in constitutive aiiA generator receiver : Went too much. |
| - | #* | + | #*[[Image:Chiba_check.png]] Tested the GFP expression in AHL-induced aiiA generator receiver : Needed to twist the circuit more. |
| - | #* | + | #*[[Image:Chiba_nocheck.png]] AHL-induced inverter aiiA receiver. Not yet assembled. |
| + | |||
===Making Marimo=== | ===Making Marimo=== | ||
Revision as of 04:14, 27 October 2007
|
Introduction | Project Design ( 1.Affinity Tag | 2.Communication Module | 3.Size Control ) | Making Marimos | Our Goal |
Project Overview
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...)
Our marimo bacteria requres as follows:
こまかすぎか．
Experiments Overview
Making Affinity Tags
Fig. The short peptide with six histidine (“His-Tag”) was inserted into the fliC D3 domain.
- 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
Making Communication Module
- Making Receivers
- AHL > GFP generator (constitutive aiiA) : BBa_T729006
- AHL > GFP & aiiA generator : BBa_I729005
- Sensitive AHL > GFP generator : BBa_I729004
inverter-aiiA
- Making Senders
- MetK Sender ( could not deposit to registry )
Controlling Size
- Improve Sender
- Overexpressed the metK (the AHL precursur synthesis enzyme) in the hope to increase AHL synthesis. (Turned out not to work)
- Improve Receiver
- Inserted 2 mutations in LuxR which is known from the paper to increase activity.
- Localize AHL
- Tested the GFP expression in constitutive aiiA generator receiver : Went too much.
- Tested the GFP expression in AHL-induced aiiA generator receiver : Needed to twist the circuit more.
- AHL-induced inverter aiiA receiver. Not yet assembled.
