IGEM:IMPERIAL/2006/project/Oscillator/project browser/Test Sensing Predator Construct/Design: Difference between revisions

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=Predator Test Construct=
*Two parts were created to fulfil the specifications for the predator sensing test construct
These two parts are very similar and have the same inputs and outputs, therefore they can be analysed in exactly the same way


These designs should meet the specifications mentioned above and operate as shown in fig1


'''fig1'''
::*a monocystronic <bbpart>J37020</bbpart>
[[image:AHL_GFP.JPG ]]
[[image:Monocistronic.JPG ]]


The differences are in the rates of production of GFP to LuxR. Part J37016 will produce a polycistronic strand of mRNA and the rate of GFP to LuxR translation will depend on which RBS is farthest upstream. The second design; Part J37020 produces two strands of mRNA they are produced by the same promoter, LuxpR but the seuqance surrounding luxpR differs between the two sites so there may be differing rates of transcription between the promoters controlling GFP and luxR. A more reliable method to get the luxR rates of expression would be to tag LuxR with an antigen then run an assay for it. This would however require a very large amount o lab work and the modls do not need luxR translation, simply the transfer function of HSL -> aiiA


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*<big>'''[http://parts.mit.edu/registry/index.php/Part:BBa_J37016 J37016]'''</big>
<br>


::*a polycystronic <bbpart>J37016</bbpart>
[[image:polycistronic.JPG]]
[[image:polycistronic.JPG]]




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*<big>'''[http://parts.mit.edu/registry/index.php/Part:BBa_J37020  J37020]'''</big>
<br>
[[image:Monocistronic.JPG ]]


*These two parts are very similar and have the same inputs and outputs, therefore they can be analysed in exactly the same way
The differences are in the rates of production of GFP to LuxR:
*Part J37016 will produce a polycistronic strand of mRNA and the rate of GFP to LuxR translation will depend on which RBS is farthest upstream.
*Part J37020 produces two strands of mRNA they are produced by the same promoter, LuxpR but the seuqance surrounding luxpR differs between the two sites so there may be differing rates of transcription between the promoters controlling GFP and luxR.


==Design choices==
==Design choices==
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! OUTPUTS !! biological component !! comments
! OUTPUTS !! biological component !! comments
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| Green Flourescent Protein || GFP || GFP is synthesed by the <bbpart>BBa_E0040</bbpart> gene
| Green Flourescent Protein || GFP || GFP is synthesed by the <bbpart>BBa_E0040</bbpart> gene. This gene is in place of AiiA in the predator construct. We make the assumption that rate of production of GFP is equal to the rate of production of AHL-lactonase to generate a transfer curve to allow characterization of the predator sensing part.
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|- style="background:lightgrey"
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Latest revision as of 07:37, 29 October 2006

Super Parts Predator Construct
Actual Part
Sub Parts <bbpart>R0062</bbpart> <bbpart>B0034</bbpart> <bbpart>C0062</bbpart> <bbpart>I13504</bbpart>


  • Two parts were created to fulfil the specifications for the predator sensing test construct


  • a monocystronic <bbpart>J37020</bbpart>


  • a polycystronic <bbpart>J37016</bbpart>


  • These two parts are very similar and have the same inputs and outputs, therefore they can be analysed in exactly the same way


The differences are in the rates of production of GFP to LuxR:

  • Part J37016 will produce a polycistronic strand of mRNA and the rate of GFP to LuxR translation will depend on which RBS is farthest upstream.
  • Part J37020 produces two strands of mRNA they are produced by the same promoter, LuxpR but the seuqance surrounding luxpR differs between the two sites so there may be differing rates of transcription between the promoters controlling GFP and luxR.

Design choices

  • We need to fulfill all the properties listed in the specifications.
  • Based on the Quorum-sensing/quenching due to biochemical properties needed and BioBricks availability.


INPUTS biological component comments
Prey molecule AHL Acyl Homoserine lactone (AHL) is synthesized by the LuxI gene <bbpart>BBa_C0061</bbpart>
OUTPUTS biological component comments
Green Flourescent Protein GFP GFP is synthesed by the <bbpart>BBa_E0040</bbpart> gene. This gene is in place of AiiA in the predator construct. We make the assumption that rate of production of GFP is equal to the rate of production of AHL-lactonase to generate a transfer curve to allow characterization of the predator sensing part.
FUNCTIONS biological component comments
Prey Sensing pLux promoter <bbpart>BBa_R0062</bbpart> The pLux promoter allows us to get the growth of the predator to be function of the population of prey-molecules and predator-molecules.