From 2006.igem.org
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| *[http://www3.imperial.ac.uk/people/d.mann Dr. David Mann] | | *[http://www3.imperial.ac.uk/people/d.mann Dr. David Mann] |
| *Kirsten Jensen | | *Kirsten Jensen |
- | *[http://openwetware.org/wiki/User:Vincent| Vincent Rouilly] | + | *[http://openwetware.org/wiki/User:Vincent Vincent Rouilly] |
- | *[http://openwetware.org/wiki/User:chuehloo| Chueh-Loo Poh] | + | *[http://openwetware.org/wiki/User:chuehloo Chueh-Loo Poh] |
- | *[http://openwetware.org/wiki/User:SxE00| Matthieu Bultelle] | + | *[http://openwetware.org/wiki/User:SxE00 Matthieu Bultelle] |
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Revision as of 01:43, 30 October 2006
Imperial College London iGEM 2006
[http://openwetware.org/wiki/IGEM:IMPERIAL/2006 Full Documentation of the project on OWW]
| Project Summary:
Oscillators are a fundamental building block in many engineering fields and are a widespread phenomenon in biology. Building a biological oscillator is thus a crucial step forward in the field of Synthetic Biology.
Engineering a Molecular Predation Oscillator, the iGEM project 2006 of Imperial College London, provides a new approach to create a stable biological oscillator: It follows an engineering-based cycle of specification, design, modelling, implementation and testing/validation. The innovative design of the oscillator relies on predator-prey dynamics based on the Lotka-Volterra model.
| Achievements:
Following the design of the oscillator, a full theoretical analysis of the Lotka-Volterra properties was carried out, which promised the successful outcome of oscillations. Additionally, all components as well as the overall oscillator were modelled such that the behaviour of the system could be predicted. Our team successfully built functional parts, thus providing the building block for the final oscillator. All parts created were experimentally tested and their characterization could be used to feedback information into the modelling.
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Main Project
| Our Approach: Engineering a Molecular Predation Oscillator
For building our biological oscillator, the engineering cycle below was followed. Click on each of the stages of the engineering cycle below in order to find out about the different stages & aspects of the project.
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1) Specifications:
- Building a stable biological oscillator with controllable oscillations in a modular design
2) Design:
- Mimics predator-prey dynamics based on the Lotka-Volterra model
3) Modelling:
- Full theoretical analysis
- Modelling of the full system
4) Implementation:
- 4 functional & 5 intermediate parts successfully submitted to the Registry
5) Testing/Validation:
- Testing of different components of the oscillator, thus characterizing the parts
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Secondary Projects
[http://openwetware.org/wiki/IGEM:IMPERIAL/2006/project/popsblocker PoPs Blocker] | [http://openwetware.org/wiki/IGEM:IMPERIAL/2006/project/Bio_elec_interface Biological to Electrical Interface]
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As a method of controlling the activation of the positive-feedback loop in our predator-prey based oscillator, we successfully created this part, which can be used as a general Pops Blocker:
- This part is placed downstream of a promoter and prevents any Pops from the promoter passing through this part
- When an accompanying Cre Recombinase plasmid becomes activated, the enzyme produced will permanently cut a section of DNA from the plasmid containing this part
- Only then, the polymerase can pass through this part and transcribe downstream genes.
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We also worked on a Biosensor for measuring AHL concentrations in order to establish a Biological to Electrical Interface this summer.
- Using an enzyme to hydrolize the lactone AHL would result in a local change in pH
- Measuring the change in pH gives a measurement of how much AHL is present
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Our Contributions to the Registry
Part Logo | Description | Link to registry | Built | Tested | Characterized | Sequenced | Sent
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Final Constructs
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| Final Prey Cell | J37015 | YES | YES | Pending | YES | YES
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| Cre/Lox Prey Control | J37027 | YES | YES | YES | YES | YES
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Test Constructs
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| Final Polycistronic Predator Cell Test Construct | J37016 | YES | YES | YES | YES | YES
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| Predator Cell, pLux Transfer Function (two promoters) | J37020 | YES | YES | NO | NO | YES
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Intermediate Parts
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| AHL induced LuxR generator (for predator cell) | J37019 | YES | N/A | N/A | YES | YES
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| RBS + LuxR | J37033 | YES | N/A | N/A | YES | YES
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| Prey Cell Intermediate | J37034 | YES | N/A | N/A | YES | YES
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| LuxR + GFP | J37032 | YES | N/A | N/A | YES | YES
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Our Open Documentation
- [http://openwetware.org/wiki/IGEM:IMPERIAL/2006/LabCalendar| Lab Notebook]
- [http://openwetware.org/wiki/IGEM:IMPERIAL/2006/Protocols| Protocols]
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- [http://openwetware.org/wiki/IGEM:IMPERIAL/2006/Journal Club| Journal Club]
- [http://openwetware.org/wiki/IGEM:IMPERIAL/2006/Lecture Notes| Lecture Notes]
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- [http://openwetware.org/wiki/BioSysBio:abstracts/2007/iGEM2006_Imperial_College BioSysBio Conference, UK]
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- [http://openwetware.org/wiki/IGEM:IMPERIAL/2006/Calendar/2006-7-26| UK iGEM Teams Meeting]
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- [http://openwetware.org/wiki/IGEM:IMPERIAL/2006/Photos| Photos]
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The Team and Acknowledgements
Undergrads
- [http://openwetware.org/wiki/User:Cys Christin Sander]
- [http://openwetware.org/wiki/User:da104 Deepti Aswani]
- [http://openwetware.org/wiki/User:Farah Farah Vohra]
- [http://openwetware.org/wiki/User:Baijiongjun Jiongjun Bai]
- [http://openwetware.org/wiki/User:Johnsy John Sy]
- [http://openwetware.org/wiki/User:JohnChattaway John Chattaway]
- [http://openwetware.org/wiki/User:Jonny Jonathan Wells]
- [http://openwetware.org/wiki/User:Tom Tom Hinson]
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The Imperial College iGEM team and advisors on the terrace of the Biochemistry building
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Advisors
- [http://www3.imperial.ac.uk/people/r.kitney Prof. Richard Kitney]
- [http://www3.imperial.ac.uk/people/p.freemont Prof. Paul Freemont]
- [http://www3.imperial.ac.uk/people/d.mann Dr. David Mann]
- Kirsten Jensen
- [http://openwetware.org/wiki/User:Vincent Vincent Rouilly]
- [http://openwetware.org/wiki/User:chuehloo Chueh-Loo Poh]
- [http://openwetware.org/wiki/User:SxE00 Matthieu Bultelle]
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Acknowledgements
- Prof. Tony Cass
- Dr. Anna Radomska
- Dr. Rupert Fray
- Dr. David Leak
- Dr. Mauricio Barahona
- Dr. Danny O'Hare
- Dr. Geoff Baldwin
- Dr. Ian Smith
- Susan E. Wryter
- David Featherbe
- Ciaran Mckeown
- James Mansfield
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Funding
- European Commission
- Imperial College Deputy Rector's Fund
- Imperial College Faculty of Engineering
- Imperial College Faculty of Natural Sciences
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