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Team:Chiba/Sender experiments
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===Method=== | ===Method=== | ||
| + | 1.Crosstalk test | ||
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To characterize quorum sensing crosstalk, constitutive AHL senders were mixed with constitutive receivers and measure fluorescence intensity. | To characterize quorum sensing crosstalk, constitutive AHL senders were mixed with constitutive receivers and measure fluorescence intensity. | ||
| - | #Transformed Senders into E.coli strains( | + | #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 | #Inoculated them independently in liquid media. Incubated at 37°C 12h | ||
#Mixed them. | #Mixed them. | ||
#Incubated at 30°C. | #Incubated at 30°C. | ||
#Measured intensity of green fluorescence at regular time intervals. | #Measured intensity of green fluorescence at regular time intervals. | ||
| + | |||
| + | 2.Demonstration | ||
| + | |||
===Result=== | ===Result=== | ||
Revision as of 01:02, 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
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
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
1.Crosstalk test
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.
- 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）
2.Demonstration
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