Team:Chiba/Sender experiments
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[[image:senders_crosstalk_chiba_01.gif|frame|left| | [[image:senders_crosstalk_chiba_01.gif|frame|left| | ||
'''Fig.''' senders crosstalk test.senders strain XL10Gold,Receiver strain JW1908.Reaction temparature was 30°C.All measurements are averages from three replicate cultures with error bars representing standard deviations.Labeling:LuxI,RhlI,LasI means fluorescence induced by AHLs synthesized by LuxI,RhlI,LasI respectively.]]<br clear="all"> | '''Fig.''' senders crosstalk test.senders strain XL10Gold,Receiver strain JW1908.Reaction temparature was 30°C.All measurements are averages from three replicate cultures with error bars representing standard deviations.Labeling:LuxI,RhlI,LasI means fluorescence induced by AHLs synthesized by LuxI,RhlI,LasI respectively.]]<br clear="all"> | ||
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(冨永) | (冨永) | ||
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Revision as of 01:03, 26 October 2008
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Contents |
Quorum-Sensing Cross-talk
Purpose
To Confirm that communication using non-endogenous molecules results in slower activation of receivers.
冨永
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
1.Crosstalk test
To characterize quorum sensing crosstalk, constitutive AHL senders were mixed with constitutive receivers and measure fluorescence intensity.
- Transformed Senders into E.coli strains(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.
2.Demonstration
Result
Crosstalk test
- 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.
(冨永)
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