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Team:Chiba/Sender experiments
From 2008.igem.org
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Quorum-Sensing Cross-talk
Purpose
To Confirm that communication using non-endogenous molecules results in slower activation of receivers.
冨永
Fig.5 Senders crosstalk
Senders
- [http://partsregistry.org/Part:BBa_K084007 plac+rbs+LasI]
- [http://partsregistry.org/Part:BBa_K084008 plac+rbs+RhlI]
- [http://partsregistry.org/Part:BBa_K084012 plac+rbs+LuxI]
Receiver
- [http://partsregistry.org/Part:BBa_T9002 BBa_T9002 (Express GFP in response to AHL)]
Method
To characterize quorum sensing crosstalk, constitutive AHL senders were mixed with constitutive receivers and measure fluorescence intensity.
- Transformed Senders into E.coli strains(JW1908/XL10GOLD) and Receiver into E.coli strain(JW1908).
- Inoculated them independently in liquid media. Incubated at 37°C 12h
- Mixed them.
- Incubated at 30°C.
- Measured intensity of green fluorescence at regular time intervals.
Result
1.Crosstalk test at 30°C
Fig. senders crosstalk test.senders strain XL10Gold,Receiver strain JW1908.Reaction temparature was 30°C.Labeling:LuxI,RhlI,LasI means fluorescence induced by AHLs synthesized by LuxI,RhlI,LasI respectively.
- RhlIとLuxIでは、GFP inductionにかかる時間はほとんど同じであった。
- LasIは、GFP inductionが他より約2時間遅れた。
2.Demonstration
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