IGEM:IMPERIAL/2007/Projects/Biofilm Detector/Design: Difference between revisions

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====GFP Reporter====
====GFP Reporter====
[http://parts.mit.edu/registry/index.php/Part:BBa_E0040 BBa_E0040] GFP<br>
[http://parts.mit.edu/registry/index.php/Part:BBa_I13504 BBa_EI1354] GFP<br>
Standard GFP, excited/emitted at 501/511 nm
Standard GFP, excited/emitted at 501/511 nm, with RBS and stop codons.


==Stage 1: Detector for AHL==
==Stage 1: Detector for AHL==

Revision as of 11:52, 29 July 2007

Biofilm Detection: Design



Overall Design


Possible Parts

Signaling System

BBa_F1610 RBS-luxI
No promoter - we need to put one on it!

Promoter for Signaling System

BBa_R0040 ptet, if uncontrolled
pLac, if constitutive secretion
pBAD, if controlled secretion

Detection System

BBa_F2620 ptet-LuxR-pLux
Detection range: 1-1000 nM, highest sensitivity from 1-10 nM
Peak secretion: 400 GFP molecules cfu-1 s-1
Peak response time: 27 minutes

GFP Reporter

BBa_EI1354 GFP
Standard GFP, excited/emitted at 501/511 nm, with RBS and stop codons.

Stage 1: Detector for AHL

Our first aim is to create a system that can accurately detect the concentration of AHL in solution. We found a well characterized detector system sensitive to 3OC6HSL which we will attach to a standard GFP reporter to determine its response to GFP.
 

Stage 2: Detector for bacterial secreted AHL

To more accurately simulate a bacterial biofilm, we decided to create bacteria that would constitutively secrete AHL.