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.

の培養液を混ぜた反応液は、GFPの蛍光強度が上昇しなかった。BBa_K084010が働いていないか、クロストークが起こっていないことが考えられる。<-- DON'T DELETE -->

  1. Others responded at the same time(4 hours after induction).Final fluorescence intensity differd.

CinI+LVA以外の3種の反応液は,すべて立ち上がりは同じであり(4時間後),LuxI,RhlIとLasIとで,最終到達蛍光強度に差が生じた.<-- DON'T DELETE -->

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.

立ち上がりの時間,最終蛍光強度ともに,ほとんど差が見られなかった.induction後,AHL濃度はすぐに閾値に達しており,どの反応液もgfpが成熟するのに伴い,蛍光強度が上昇していると考えた.

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.

LVAtagの効果は見られなかった.LVAによってシンターゼが分解されるよりも速い速度でAHLが合成されていると考えた.

Reaction temparature:30°C

センダーの培養液：500μL、レシーバの培養液：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.
  2. LVAtagの効果は見られなかった.LVAによってシンターゼが分解されるよりも速い速度でAHLが合成されていると考えた.

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


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.
  2. LVAtagの効果は見られなかった.LVAによってシンターゼが分解されるよりも速い速度でAHLが合成されていると考えた.

Sender culture：10μL,Receiver culture：1000μL

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


  1. 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 increased.

LVAtagがついている場合,ついていないものより最終到達蛍光強度が小さくなった.LVAによってシンターゼが分解されていた.しかし,立ち上がりの時間は変化しなかった.これは,シンターゼよりAHL合成の速度が速いために,AHLがすぐに閾値に達してしまっているためと考えた.閾値を超えると,すぐさまgfpの翻訳が始まり,inductionの2時間後には蛍光強度が上昇し始めた.


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