Team:Chiba/Receiver experiments



Design
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）

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 ReceiverBBa_T9002 (AHL Reporter)




 * Medium Copy Receiver



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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