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Team:Chiba/Sender experiments
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Signal molecule sender
Design
Each species has their own LuxI-type proteins,which synthesize their specific autoinducers,AHLs.AHLs produced by different LuxI-type proteins differ only in the length of the acyl-chain moiety and substitution at position C-3.LuxR,which is original for Vibrio fischeri,is activated by its cognate autoinducer,3OC6HSL.However,LuxR is also activated by non-endogenous molecules,C4HSL,C6HSL,and 3OC12HSL.Activation by non-endogenous molecules requires a higher signal concentration(2).This results in slower activation of receivers,when AHL concentration is increasing.
Experiment
To Confirm that communication using non-endogenous molecules results in slower activation of receivers.
冨永
Senders
Receiver
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.
Result
Crosstalk test
- 軸ラベル、直します。
- 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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