IGEM:IMPERIAL/2006/project/Oscillator/project browser/Prey Construct/Design

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*The prey part: [http://parts.mit.edu/registry/index.php/Part:BBa_J37015 J37015 @ MIT Parts Registry]
*The prey part: [http://parts.mit.edu/registry/index.php/Part:BBa_J37015 J37015 @ MIT Parts Registry]
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[[Image:J37015 logo.png]]
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[[Image:J37015page.JPG]]
==Design Choices==
==Design Choices==
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*Design allows for an exponential increase of AHL via a positive feedback loop
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*Design must fulfil the [http://openwetware.org/wiki/IGEM:IMPERIAL/2006/project/Oscillator/project_browser/Prey_Construct specifications].
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**Interaction between LuxR, AHL and promoter controlling expression of LuxI
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*Part design will be based on Quorum-sensing/quenching and the availability of BioBricks.
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::*Research into the registry directed us to use AHL as our prey molecule.
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*Design allows for an exponential increase of AHL (to fulfil the [[IGEM:IMPERIAL/2006/project/Oscillator/Theoretical_Analyses/2D_Model1|Lotka-Volterra model]]) via a positive feedback loop.
 +
*The prey part can be decoupled into a prey sensing part and a prey producing part.
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*Initiation of positive feedback loop depends on leaky expression of the tetR promoter which is a constitutive promoter.
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** This leaky expression produces LuxR which can form a dimer with AHL already withing the cell.
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** LuxR, AHL and promoter control the expression of LuxI which produces AHL.
*Design allows for assesment of status of positive feedback loop via:
*Design allows for assesment of status of positive feedback loop via:
**GFP Production - Linked to LuxI expression
**GFP Production - Linked to LuxI expression
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**AHL Production - Can be indirectly assessed via T9002 assay
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**AHL Production - Can be indirectly assessed according to [http://openwetware.org/wiki/IGEM:IMPERIAL/2006/Protocols/J37015 J37015 protocol].
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! INPUTS !! biological component !! comments
! INPUTS !! biological component !! comments
|-
|-
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|AHL||LuxR + pLuxR||These components sense input and activate output
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|AHL||LuxR & pLuxR|| To initiate the positive feedback loop, LuxR is produced via leaky expression of the tetR promoter.
|- style="background:orange"
|- style="background:orange"
! OUTPUTS !! biological component !! comments
! OUTPUTS !! biological component !! comments
|-
|-
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|AHL||LuxI||LuxI expression causes enzymatic generation of AHL  
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|AHL||LuxI||LuxR, AHL and pLuxR form a complex, activating production of LuxI expression which causes enzymatic generation of AHL  
|}
|}

Revision as of 21:02, 29 October 2006

Super Parts Molecular Prey-Predator Oscillator
Actual Part Image:J37015page.JPG
Sub Parts Test Sensing Prey Construct <bbpart>C0261</bbpart> <bbpart>I13401</bbpart>


Registry

Image:J37015page.JPG

Design Choices

  • Design must fulfil the specifications.
  • Part design will be based on Quorum-sensing/quenching and the availability of BioBricks.
  • Research into the registry directed us to use AHL as our prey molecule.
  • Design allows for an exponential increase of AHL (to fulfil the Lotka-Volterra model) via a positive feedback loop.
  • The prey part can be decoupled into a prey sensing part and a prey producing part.
  • Initiation of positive feedback loop depends on leaky expression of the tetR promoter which is a constitutive promoter.
    • This leaky expression produces LuxR which can form a dimer with AHL already withing the cell.
    • LuxR, AHL and promoter control the expression of LuxI which produces AHL.
  • Design allows for assesment of status of positive feedback loop via:
    • GFP Production - Linked to LuxI expression
    • AHL Production - Can be indirectly assessed according to J37015 protocol.


INPUTS biological component comments
AHLLuxR & pLuxR To initiate the positive feedback loop, LuxR is produced via leaky expression of the tetR promoter.
OUTPUTS biological component comments
AHLLuxILuxR, AHL and pLuxR form a complex, activating production of LuxI expression which causes enzymatic generation of AHL
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