IGEM:IMPERIAL/2007/Projects/Biofilm Detector/Design: Difference between revisions
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[http://parts.mit.edu/registry/index.php/Part:BBa_I13504 BBa_EI1354] GFP<br> | [http://parts.mit.edu/registry/index.php/Part:BBa_I13504 BBa_EI1354] GFP<br> | ||
Standard GFP, excited/emitted at 501/511 nm, with RBS and stop codons.<br> | Standard GFP, excited/emitted at 501/511 nm, with RBS and stop codons.<br> | ||
LacZ for β-galactosidase assay | LacZ for β-galactosidase assay (yet to be discovered...) | ||
==Stage 1: Detector for AHL== | ==Stage 1: Detector for AHL== | ||
Revision as of 13:21, 2 August 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
Part BBa_J24677 already has GFP attached
GFP Reporter
BBa_EI1354 GFP
Standard GFP, excited/emitted at 501/511 nm, with RBS and stop codons.
LacZ for β-galactosidase assay (yet to be discovered...)
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.
We would later test the promoter on a different reporter.
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 detector plasmid would incorporate the Hrp system as an amplifier, to amplify a reporter gene yet to be decided.
