Chiba/Making Marimo

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*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.
*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====
====Results====
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Failed to ligate FliC-His generator and p15A origin vector.<br>
+
First attempt to import FliC-His generator into p15A vector (see Fig).<br>
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*混乱しています。ごめん。確認なんですが、FliC-His generatorをp15Aにのせるというのは、Biobrickにするためでないでしょうか。。僕の勘違いかな？。。そもそもQuorum sensing のベクターと、FliC-Hisチェックしてビーズ吸着させたときのベクターとを、ダブルトランスフォーメーションできないんだっけ？byとよたろ
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Communication units are on ColE1 type plasmids. <br>
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To make this ''stickly FliC'' construct compatible with communication circuits, we need to do this.
===[[Chiba/Flagella/FliC-His_Biobrick|FliC-his biobrick]]===
===[[Chiba/Flagella/FliC-His_Biobrick|FliC-his biobrick]]===

Revision as of 04:18, 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

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, we need to do this.

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). Cloning is still underway -->

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