Team:Chiba/Sender experiments

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Quorum-Sensing Cross-talk

Purpose

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

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

  1. Transformed Senders into E.coli strains(JW1908/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.

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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