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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)]
- 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。
- 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).
- Inoculated them independently in liquid media. Incubated at 37℃ 12h.
- Inoculated again in Fresh liquid media upto about OD600=2 at 37℃
- Washed sender and receivers.
- Mixed them. (Sender:Receiver=1000μL:1000μL)
- Incubated at 30°C.
- Measured intensity of green fluorescence at regular time intervals.(Fluoroskan AscentR FL＆Fluoroskan AscentR Thermo ELECTRON CORPORATION)
Result
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