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

From 2008.igem.org

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.

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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