Team:Chiba/Sender experiments/Senders(JW1908) T9002(JW1908)

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Results and Discussion

Reaction temparature:37°C,09/12

Sender culture:1000μL,Receiver culture:1000μL

Fig.1  
12/09.E.coli strain,senders:JW1908,BBa_T9002:JW1908,Reaction temparature:37°C,Receiver cells/Sender cells = 1.


Sender culture:100μL,Receiver culture:1000μL

Fig.2  
E.coli strain,senders:JW1908,BBa_T9002:JW1908,37°C,Receiver cells/Sender cells = 10.



  1. Green fluorescence intensity didn't increase in the culture containing BBa_K0840010(plac+CinI+LVA).We thought BBa_K084010 didn't work properly or LuxR gene didn't interact with signal molecules synthesized by BBa_K084010.
  1. Others responded at the same time(4 hours after induction).Final fluorescence intensity differd.


Reaction temparature:37°C

Sender culture:500μLm,Receiver culture:500μL

Fig.3  
E.coli strain,senders:JW1908,BBa_T9002:JW1908,37°C
Fig.4  
E.coli strain,senders:JW1908,BBa_T9002:JW1908,37°C


Left:

  1. Response time and final fluorescence intensity showed no significant difference.

We thought AHL concentration was quickly reached the threshold concentration.The fluorescence intensity increased as gfp maturured.

Right:

  1. No significant difference in fluorescence intensity between the culture containing BBa_K084007(plac+RhlI) gene transformed cells and the culture containing BBa_K084008(plac+RhlI(LVA)) gene transformed cells.We thought that the rate of AHL synthesis by each autoinducer synthase was much faster than the rate of its degradation by protease.

Reaction temparature:30°C

Sender culture:500μL,Receiver culture:500μL

Fig.5  
E.coli strain,Senders:JW1908,BBa_T9002:JW1908,30°C,Receiver cells/Sender cells = 1.
Fig.6  
E.coli strain,Senders:JW1908,BBa_T9002:JW1908,30°C,Receiver cells/Sender cells = 1.


Left:

Right:

  1. No significant difference in fluorescence intensity between the culture containing BBa_K084007(plac+RhlI) gene transformed cells and the culture containing BBa_K084008(plac+RhlI(LVA)) gene transformed cells.We thought that the rate of AHL synthesis by each autoinducer synthase was much faster than the rate of its degradation by protease.


Sender culture:100μL,Receiver culture:1000μL

Fig.7  
E.coli strain,senders:JW1908,BBa_T9002:JW1908,30°C
Fig.8 
E.coli strain,senders:JW1908,BBa_T9002:JW1908,30°C


No significant difference in fluorescence intensity between the culture containing BBa_K084007(plac+RhlI) gene transformed cells and the culture containing BBa_K084008(plac+RhlI(LVA)) gene transformed cells.We thought that the rate of AHL synthesis by each autoinducer synthase was much faster than the rate of its degradation by protease.

Sender culture:10μL,Receiver culture:1000μL

Fig.9  
E.coli strain,senders:JW1908,BBa_T9002:JW1908,30°C,Receiver cells/Sender cells = 100.


The final fluorescence intensity of the culture containing BBa_K084008 transformed cells which express LVA-tagged RhlI protein was lower than the culture containing cells which express untagged RhlI protein.It shows that LVA-tagged RhlI protein was degraded by LVA-specific protase.However,fluorescence intensity of both cultures increased at the same time.We thought it was because the AHL concentration was quickly reached the threshold concentration and cells began to express gfp.After 2 hours,fluorescence intensity started to increase.


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