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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
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
E.coli strain,senders:JW1908,BBa_T9002:JW1908,37°C,Receiver cells/Sender cells = 10.
- 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 -->
- 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
E.coli strain,senders:JW1908,BBa_T9002:JW1908,37°C
E.coli strain,senders:JW1908,BBa_T9002:JW1908,37°C
Left:
- 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:
- 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
E.coli strain,Senders:JW1908,BBa_T9002:JW1908,30°C,Receiver cells/Sender cells = 1.
E.coli strain,Senders:JW1908,BBa_T9002:JW1908,30°C,Receiver cells/Sender cells = 1.
Left:
Right:
- 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.
Sender culture：100μL,Receiver culture：1000μL
E.coli strain,senders:JW1908,BBa_T9002:JW1908,30°C
E.coli strain,senders:JW1908,BBa_T9002:JW1908,30°C
Left:
Right:
- 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.
Sender culture：10μL,Receiver culture：1000μL
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
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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