Team:Chiba/Sender experiments

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(Difference between revisions)
(1.Crosstalk test)
(Quorum-Sensing Cross-talk)
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===Method===
===Method===
 +
1.Crosstalk test
 +
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.
   
   
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#Transformed Senders into E.coli strains(JW1908/XL10GOLD) and Receiver into E.coli strain(JW1908).
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#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

Chiba-U.gif

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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]

Tet-lasi chiba.gif

  • [http://partsregistry.org/Part:BBa_K084008 plac+rbs+RhlI]

Tet-rhli chiba.gif

  • [http://partsregistry.org/Part:BBa_K084012 plac+rbs+LuxI]

Tet-luxi chiba.gif

Receiver

  • [http://partsregistry.org/Part:BBa_T9002 BBa_T9002 (Express GFP in response to AHL)]

Reporter 9002 chiba.gif

Method

1.Crosstalk test

To characterize quorum sensing crosstalk, constitutive AHL senders were mixed with constitutive receivers and measure fluorescence intensity.

  1. Transformed Senders into E.coli strains(XL10GOLD) and Receiver into E.coli strain(JW1908).
  2. Inoculated them independently in liquid media. Incubated at 37°C 12h
  3. Mixed them.
  4. Incubated at 30°C.
  5. 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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