IGEM:IMPERIAL/2007/Projects/Biofilm Detector/Design: Difference between revisions
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==Stage 2: Detector for bacterial secreted AHL== | ==Stage 2: Detector for bacterial secreted AHL== | ||
To more accurately simulate a bacterial biofilm, we decided to create bacteria that would constitutively secrete AHL.<br> | To more accurately simulate a bacterial biofilm, we decided to create bacteria that would constitutively secrete AHL.<br> | ||
[[Image:IC2007_BF_stage2_construct_sender.png]]<br clear="all> | [[Image:IC2007_BF_stage2_construct_sender.png]]<br clear="all> <br> | ||
Our sender plasmid would incorporate the Hrp system as an amplifier, to amplify a reporter gene yet to be decided.<br> | |||
[[Image:IC2007_BF_stage2_construct_receiver.png]]<br clear="all> <br> | |||
Revision as of 19:48, 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.
Our sender plasmid would incorporate the Hrp system as an amplifier, to amplify a reporter gene yet to be decided.
