Chiba/Making Marimo
From 2007.igem.org
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Introduction | Project Design ( 1.Affinity Tag | 2.Communication Module | 3.Size Control ) | Making Marimos | Our Goal |
Making Marimos
Fig1. Scheme of final gene cuircuit for Bacteria Marimo.
Parts Construction
Because we found that FliC code includes restriction enzyme(EcoRI,SpeI,PstI) which is used in Biobrick, we divided into plasmid of FliC and one of signal, and aimed the double transformation.
FliC-His generator
Experiment
Fig2.Ligation Strategy
- We regulate His-tagged FliC by lux promoter. Namely, if LuxR is expressed, bacteria can express FliC by Quorum Seinsing.
- Because the gene circuit of Quorum Sensing is on vector of colEI, we can use FliC and Quorum Sensing by using vector of p15A and do double transformation.
Results
First attempt to import FliC-His generator into p15A vector (see Fig).
Communication units are on ColE1 type plasmids.
To make this stickly FliC construct compatible with communication circuits, this is an absolute necessity.....
FliC-his biobrick
Experiment
- Making Biobrick version of Flic-His and other stickly-FliCs. unfortunately, FliC has many EcoRI,SpeI,PstI sites. We have to eliminate them one by one by site-directed mutagenesis.
Results
Not yet (as of 10/26/2007). The first ligation sucked.
Cloning is still underway -->
