Cambridge

From 2007.igem.org

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In order to engineer interesting and useful functions in biology, a robust and extensive range of intra- and inter-cellular signalling pathways must be available. By analogy with the Internet, where adoption of the standard TCP/IP communication protocol has enabled worldwide connectivity from supercomputers to refrigerators, such a system must be accessible to cells of different heritage and structure (different “operating systems”) with the potential for processing messages received and taking action dependent on their content (see diagram at right).  
In order to engineer interesting and useful functions in biology, a robust and extensive range of intra- and inter-cellular signalling pathways must be available. By analogy with the Internet, where adoption of the standard TCP/IP communication protocol has enabled worldwide connectivity from supercomputers to refrigerators, such a system must be accessible to cells of different heritage and structure (different “operating systems”) with the potential for processing messages received and taking action dependent on their content (see diagram at right).  
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In the course of this project we attempted to implement candidates for both types of signalling mechanism, and additionally made progress towards adding a new Gram-positive platform for synthetic biology to the Registry.
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In the course of our project we identified and worked on candidates for both intracellular ([[Cambridge/Amplifier project|PoPS Amplifier project]]) and intercellular ([[Cambridge/Signalling project|Peptide signalling project]]) signalling pathways, and additionally made progress towards adding a new [[Cambridge/Gram-positive chassis project|Gram-positive platform for synthetic biology]] to the Registry.
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*'''Intracellular signalling:''' The <u>PoPS Amplifier Project</u>
 
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*'''Intercellular signalling:''' subproject 3).
 
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*[http://www.ccbi.cam.ac.uk/iGEM2007/index.php/Main_Page Go to the Cambridge iGEM 2007 team's official wiki!] (with more information about the team and our progress towards this goal)
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*[http://www.ccbi.cam.ac.uk/iGEM2007/index.php/Main_Page Go to the Cambridge iGEM 2007 team's official wiki!] (with more information about the team and how we came up with this idea)
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*[http://partsregistry.org/cgi/partsdb/pgroup.cgi?pgroup=iGEM2007&group=Cambridge BioBricks we have contributed to the Registry]
*[http://partsregistry.org/cgi/partsdb/pgroup.cgi?pgroup=iGEM2007&group=Cambridge BioBricks we have contributed to the Registry]
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[[Image:Camb amplif icon.png|50px|left]]
[[Image:Camb amplif icon.png|50px|left]]
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<span style="font-weight: bold; font-size: 120%; padding: 0 0 0 0.5em; line-height: 300%;">PoPS amplifier</span>
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<span style="font-weight: bold; font-size: 120%; padding: 0 0 0 0.5em; line-height: 300%;">[[Cambridge/Amplifier project|PoPS amplifier]]</span>
We BioBricked and characterised strong activators and their associated promoters. With them we built a standard amplifier which can be used in any synthetic transcriptional system, taking in a standard PoPS input and giving a PoPS output of a known amplification factor. In addition, the activator-promoter pairs studied have differential levels of activation in different combinations - this crosstalk enables their potential use for co-ordinating complex responses to stimuli.
We BioBricked and characterised strong activators and their associated promoters. With them we built a standard amplifier which can be used in any synthetic transcriptional system, taking in a standard PoPS input and giving a PoPS output of a known amplification factor. In addition, the activator-promoter pairs studied have differential levels of activation in different combinations - this crosstalk enables their potential use for co-ordinating complex responses to stimuli.
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[[Image:Camb agr icon.png|50px|left]]
[[Image:Camb agr icon.png|50px|left]]
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<span style="font-weight: bold; font-size: 120%; padding: 0 0 0 0.5em; line-height: 300%;">Peptide signalling</span>
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<span style="font-weight: bold; font-size: 120%; padding: 0 0 0 0.5em; line-height: 300%;">[[Cambridge/Signalling project|Peptide signalling]]</span>
Peptide signalling systems exist in profusion among Gram-positive bacteria and are recognised with high specificity. To implement a paradigm peptide communication channel we chose the Agr (Accessory Gene Regulator) oligopeptide-based quorum sensing mechanism from ''S. aureus''. We worked on converting it into a reusable interbacterial signalling system for communication amongst and between both Gram-positive species (''B. subtilis'') and Gram-negative bacteria (''E. coli''), based on controllable secretion and detection of the signalling peptide AIP. In order to use such hydrophilic signal molecules in Gram-negative bacteria, we introduced an outer membrane pore to allow their diffusion into the periplasm.  
Peptide signalling systems exist in profusion among Gram-positive bacteria and are recognised with high specificity. To implement a paradigm peptide communication channel we chose the Agr (Accessory Gene Regulator) oligopeptide-based quorum sensing mechanism from ''S. aureus''. We worked on converting it into a reusable interbacterial signalling system for communication amongst and between both Gram-positive species (''B. subtilis'') and Gram-negative bacteria (''E. coli''), based on controllable secretion and detection of the signalling peptide AIP. In order to use such hydrophilic signal molecules in Gram-negative bacteria, we introduced an outer membrane pore to allow their diffusion into the periplasm.  
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[[Image:Camb bacillus icon.png|50px|left]]
[[Image:Camb bacillus icon.png|50px|left]]
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<span style="font-weight: bold; font-size: 120%; padding: 0 0 0 0.5em; line-height: 300%;">Gram-positive chassis</span>
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<span style="font-weight: bold; font-size: 120%; padding: 0 0 0 0.5em; line-height: 300%;">[[Cambridge/Gram-positive chassis project|Gram-positive chassis]]</span>
Most synthetic biology so far has involved Gram-negative bacteria, which are more widely used in molecular biology research; however, the biotechnology industry relies on Gram-positive bacteria. We worked on constructing a "chassis" out of ''Bacillus subtilis'', a common Gram-positive bacterium, into which genetic engineers can add BioBrick parts and control circuits.
Most synthetic biology so far has involved Gram-negative bacteria, which are more widely used in molecular biology research; however, the biotechnology industry relies on Gram-positive bacteria. We worked on constructing a "chassis" out of ''Bacillus subtilis'', a common Gram-positive bacterium, into which genetic engineers can add BioBrick parts and control circuits.
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<span style="font-weight:bold; font-size:120%;">Other sponsors</span>
<span style="font-weight:bold; font-size:120%;">Other sponsors</span>
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* The Isaac Newton Trust-funded UROP scheme
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* The Isaac Newton Trust UROP scheme
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* EU SynBioComm programme
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* The European Union SynBioComm programme
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Revision as of 20:53, 26 October 2007

Cambridge 2007 logo.png

BOL: Bacteria OnLine
University of Cambridge iGEM 2007

Introduction

Cambridge Network diagram.png

In order to engineer interesting and useful functions in biology, a robust and extensive range of intra- and inter-cellular signalling pathways must be available. By analogy with the Internet, where adoption of the standard TCP/IP communication protocol has enabled worldwide connectivity from supercomputers to refrigerators, such a system must be accessible to cells of different heritage and structure (different “operating systems”) with the potential for processing messages received and taking action dependent on their content (see diagram at right).

In the course of our project we identified and worked on candidates for both intracellular (PoPS Amplifier project) and intercellular (Peptide signalling project) signalling pathways, and additionally made progress towards adding a new Gram-positive platform for synthetic biology to the Registry.


Camb amplif icon.png

PoPS amplifier

We BioBricked and characterised strong activators and their associated promoters. With them we built a standard amplifier which can be used in any synthetic transcriptional system, taking in a standard PoPS input and giving a PoPS output of a known amplification factor. In addition, the activator-promoter pairs studied have differential levels of activation in different combinations - this crosstalk enables their potential use for co-ordinating complex responses to stimuli.

More information on the amplifier project...

Camb agr icon.png

Peptide signalling

Peptide signalling systems exist in profusion among Gram-positive bacteria and are recognised with high specificity. To implement a paradigm peptide communication channel we chose the Agr (Accessory Gene Regulator) oligopeptide-based quorum sensing mechanism from S. aureus. We worked on converting it into a reusable interbacterial signalling system for communication amongst and between both Gram-positive species (B. subtilis) and Gram-negative bacteria (E. coli), based on controllable secretion and detection of the signalling peptide AIP. In order to use such hydrophilic signal molecules in Gram-negative bacteria, we introduced an outer membrane pore to allow their diffusion into the periplasm.

More information on the signalling project...

Camb bacillus icon.png

Gram-positive chassis

Most synthetic biology so far has involved Gram-negative bacteria, which are more widely used in molecular biology research; however, the biotechnology industry relies on Gram-positive bacteria. We worked on constructing a "chassis" out of Bacillus subtilis, a common Gram-positive bacterium, into which genetic engineers can add BioBrick parts and control circuits.

More information on the Gram-positive chassis...

Meet the team!


The Cambridge iGEM 2007 team!

Top row, from left: James Brown (PhD student mentor), Jim Haseloff (faculty supervisor), Gos Micklem (faculty supervisor)

Second row, from left: Yi Jin Liew, John Crowe (faculty), Lovelace Soirez, Stephanie May, Yue Miao

Third row, from left: Zhizhen Zhao (Jane), Stefan Milde, Dmitry Malyshev, Xinxuan Soh (Sheila)

Front row, from left: Narin Hengrung, Yi Han, David Wyatt

Not in photo: Jim Ajioka (faculty supervisor), Lorenz Wernisch (faculty), Tony Southall (postdoctoral researcher)

The mailing list for the whole Cambridge iGEM2007 team is igem2007 [at] ccbi.cam.ac.uk

Sponsors

We are extremely grateful to the following organisations for their support of our project:

https://static.igem.org/mediawiki/2007/9/95/Cambridge_UofC_coat_of_arms.png  https://static.igem.org/mediawiki/2007/1/16/Cambridge_DNA2_0_logo.png  https://static.igem.org/mediawiki/2007/f/fd/Cambridge_EPSRC_logo_small.png

Other sponsors

  • The Isaac Newton Trust UROP scheme
  • The European Union SynBioComm programme