Team:Chiba/Receiver experiments

From 2008.igem.org

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Contents

Receiver crosstalk

Design

Table.1 LuxR family gene

センダーとレシーバーを元来とは異なる組み合わせにすることによって、thresholdに達する時間を遅くする。(杉山)

Method

Result

Change Receiver's Copy Number

Design

レシーバーのコピーナンバーを減らすことで、thresholdに達するまでの時間を伸ばす。 (小林)

Method

  • High Copy Receiver[http://partsregistry.org/Part:BBa_T9002 BBa_T9002 (AHL Reporter)]

T9002.copy-number-change Chiba.gif


  • Medium Copy Receiver

Low-copy-Receiver Chiba.gif


High Copy ReceiverとMedium Copy Receiverのディレイタイムはfollowing procedureによってanalyzeした。

  1. Transformed sender(Ptet-LuxI), high copy receiver(Ptet-LuxR-Plux-GFP-colE1) and Medium copy receiver(Ptet-LuxR-Plux-GFP-p15A) respectively into E coli strains(BD⊿FliC).
  2. Inoculated them independently in liquid media. Incubated at 37℃ 12h.
  3. Inoculated again in Fresh liquid media upto about OD600=2 at 37℃
  4. Washed sender and receivers.
  5. Mixed them. (Sender:Receiver=1000μL:1000μL)
  6. Incubated at 30°C.
  7. Measured intensity of green fluorescence at regular time intervals.(Fluoroskan AscentR FL&Fluoroskan AscentR Thermo ELECTRON CORPORATION)

Result

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