Team:Chiba/jk/γ/trs

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Time Responce Solid

BBa_T9002

pLac-mCherry

shaking incubator

Innova 4200 Benchtop or Floor-Stackable Incubator Shaker(37°C)

Fig. レポーターのタイムレスポンスの実験方法

Strain:XL10G Kan^R

Pre-culture
1. Picked and cultured the following plate stocks in 2mL of LB:
  1. LB-Amp+0.2 % Glucose, (pGFPuv, pLac-Venus YFP, pLac-mCherry, pUC19)
  2. LB-Amp, ([http://partsregistry.org/Part:BBa_T9002 BBa_T9002], [http://partsregistry.org/Part:BBa_K084003 BBa_K084003])
2. Cultured at 37°C for 12h.
Spread on plate
1. Spread on new plate
  1. LB-Amp+0.2 % Glucose, (pGFPuv, pLac-Venus YFP, pLac-mCherry, pUC19)
  2. LB-Amp, ([http://partsregistry.org/Part:BBa_T9002 BBa_T9002], [http://partsregistry.org/Part:BBa_K084003 BBa_K084003])
2. Cultured at 37°C for 12h.
Colony lift
1. Colony lift to inducible agar plate (containing IPTG or AHL)
  1. LB-Amp+0.2 mM IPTG agar plate, (pGFPuv, pLac-Venus YFP, pLac-mCherry, pUC19)
  2. LB-Amp+100 nM AHL agar plate, ([http://partsregistry.org/Part:BBa_T9002 BBa_T9002], [http://partsregistry.org/Part:BBa_K084003 BBa_K084003])
2. Incubate at 37 °C
Check expression every 30 min.

Discussion Discussion

The purpose of this project is to alter the time required for expression.

expression to be observable decreased, but this still takes longer than the time required when using fluorescent proteins.

output may increase because the substrate concentration decreases with time.

For the above reasons, fluorescence proteins are more suited for our purposes.

Of the fluorescent proteins, GFP and YFP fluorescence was observable at earlier stages.

Therefore, we chose GFP as our output reporter.

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