Team:Tsinghua
From 2011.igem.org
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<h1><IMG SRC="https://static.igem.org/mediawiki/2010/d/d8/TSSLogo.png" width="40px" />Project</h1> | <h1><IMG SRC="https://static.igem.org/mediawiki/2010/d/d8/TSSLogo.png" width="40px" />Project</h1> | ||
<div class="content_block"> | <div class="content_block"> | ||
- | <div id="leftpic"><img src="https://static.igem.org/mediawiki/2010/5/5e/TSModule2m.PNG" width=90px/></div><p> | + | <div id="leftpic"><img src="https://static.igem.org/mediawiki/2010/5/5e/TSModule2m.PNG" width=90px/>'E Col Transporter' |
+ | Overview | ||
+ | This project is destined to generate several E. Coli strains which will cooperate with each other to move forth and back and transport any desired protein along a gradient. Module composition: binding, releasing, transition, movement | ||
+ | |||
+ | Binding | ||
+ | SH3 domain has high affinity for proline-rich peptides. The small size and high affinity are ideal for our carrier design. In our experiment, we used a short peptide with a Kd of 3.67μM (Y. Jacquot, et al., 2007) as the binding motif and planned to transport protein substrates with this peptide sequence. | ||
+ | This part comprises expression of three proteins, namely, OmpA-SH3 protein, which functions as the binding vehicle, OmpA-mCherry protein, which functions as the positive control, Proline-rich containing mCherry protein, which functions as the binding substrate. | ||
+ | |||
+ | Releasing | ||
+ | It’s difficult to release the substrate from strong binding, and hence protease is called into play. HIV-protease is readily available and its high efficiency, low molecular weight and high specificity constitutes fine candidate. We only need to add the substrate sequence between OmpA and SH3 sequence and the protease will cut off the binding cassette. In order to segregate the action, we’ll generate a single bacteria strain expressing the protease. | ||
+ | |||
+ | Transition | ||
+ | We need to shift between two states, the binding state and the releasing state. This molecular device is well-known for its lag in phase change, which is well adapted for our application. The lag during the phase transition is appropriate for movement and transport. | ||
+ | |||
+ | Movement | ||
+ | Kinase system is well utilized in E. Coli movement control. We’ll again use LVA tagged kinase to drive E. Coli towards the target.</div><p> | ||
<br/> | <br/> |
Revision as of 15:11, 2 July 2011
![](https://static.igem.org/mediawiki/2011/e/eb/Sh3.png)
Team
Project
Parts
![](http://biobricks.org/img/750px-BBFoundation_shadow.png)
We built a host of parts during our project. The idea is that after every small step, we store our sequence as a biobrick part.
This strategy marks our progress and facilitates future use of these sequences.
Support
Our project is supported by School of Life Sciences, Department of Physics, Academy of Arts and Designs in Tsinghua.
Besides, we cited from a series of references.
Human Practice
![](https://static.igem.org/mediawiki/2010/f/fd/Humanity.jpg)
We put safety as our first priority and made a detailed safety brochure.
We also devoted efforts to publicize synthetic biology and to cooperate with other teams. The teams in China held a summer meetup to discuss our progress and share our resources. We also held a lecture introducing iGEM and our project in Tsinghua Univeristy. To get further support, we sought the cooperation of iGEM Team at Macquire, Australia.