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

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(Sender culture:10μL,Receiver culture:1000μL)
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===Reaction temparature:37°C,09/12===
===Reaction temparature:37°C,09/12===
====Sender culture:1000μL,Receiver culture:1000μL====
====Sender culture:1000μL,Receiver culture:1000μL====
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[[Image:Chiba_talks_JW1908_37_RS1_0912_01.gif‎‎|thumb|left|'''Fig.1'''  <br>12/09.E.coli strain,senders:JW1908,BBa_T9002:JW1908,Reaction temparature:37°C,Receiver cells/Sender cells = 1.]]
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[[Image:Chiba_talks_JW1908_37_RS1_0912_01.gif‎‎|thumb|left|'''Fig.1'''  <br>12/09.E.coli strain,senders:JW1908,BBa_T9002:JW1908,Reaction temparature:37°C.All measurements are averages from three replicate cultures with error bars representing standard deviations.]]
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====Sender culture:100&mu;L,Receiver culture:1000&mu;L====
====Sender culture:100&mu;L,Receiver culture:1000&mu;L====
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[[Image:Chiba_talks_JW1908_37_RS2_0912_01.gif‎|thumb|left|'''Fig.2'''  <br>E.coli strain,senders:JW1908,BBa_T9002:JW1908,37°C,Receiver cells/Sender cells = 10.]]
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[[Image:Chiba_talks_JW1908_37_RS2_0912_01.gif‎|thumb|left|'''Fig.2'''  <br>E.coli strain,senders:JW1908,BBa_T9002:JW1908,37&deg;C.All measurements are averages from three replicate cultures with error bars representing standard deviations.]]
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#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.
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*Fig. 1,Fig. 2
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<!-- DON'T DELETE -->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 -->
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*Results
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#Others responded at the same time(4 hours after induction).Final fluorescence intensity differd.
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**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.
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<!-- DON'T DELETE -->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 -->
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**Others responded at the same time(4 hours after induction).Final fluorescence intensity differd.
===Reaction temparature:37°C===
===Reaction temparature:37°C===
====Sender culture:500&mu;Lm,Receiver culture:500&mu;L====
====Sender culture:500&mu;Lm,Receiver culture:500&mu;L====
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[[Image:Chiba_communication_JW1908_37_RS1_01.gif‎|thumb|left|'''Fig.3'''  <br>E.coli strain,senders:JW1908,BBa_T9002:JW1908,37&deg;C]]
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[[Image:Chiba_communication_JW1908_37_RS1_01.gif‎|thumb|left|'''Fig.3'''  <br>E.coli strain,senders:JW1908,BBa_T9002:JW1908,37&deg;C.All measurements are averages from three replicate cultures with error bars representing standard deviations.]]
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[[Image:Chiba_talks_JW1908_37_RS1_02.gif‎|thumb|left|'''Fig.4'''  <br>E.coli strain,senders:JW1908,BBa_T9002:JW1908,37&deg;C]]
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[[Image:Chiba_talks_JW1908_37_RS1_02.gif‎|thumb|left|'''Fig.4'''  <br>E.coli strain,senders:JW1908,BBa_T9002:JW1908,37&deg;C.All measurements are averages from three replicate cultures with error bars representing standard deviations.]]
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Left:
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*Fig. 3
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#Response time and final fluorescence intensity showed no significant difference.
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*Results
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We thought AHL concentration was quickly reached the threshold concentration.The fluorescence intensity increased as gfp maturured.
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**Response time and final fluorescence intensity showed no significant difference.
 +
*Discussion
 +
**We thought AHL concentration was quickly reached the threshold concentration.The fluorescence intensity increased as gfp maturured.
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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.
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*Fig. 4
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*Results
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Right:
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**No significant difference in fluorescence intensity between the culture containing [http://partsregistry.org/Part:BBa_K084007 BBa_K084007](plac+RhlI) gene transformed cells and the culture containing [http://partsregistry.org/Part:BBa_K084008 BBa_K084008](plac+RhlI(LVA)) gene transformed cells.
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#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.
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*Discussion
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+
**We thought that the rate of AHL synthesis by each autoinducer synthase was much faster than the rate of its degradation by protease.
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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.
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===Reaction temparature:30°C===
===Reaction temparature:30°C===
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====センダーの培養液:500&mu;L、レシーバの培養液:500&mu;L====
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====Sender culture:500&mu;L,Receiver culture:500&mu;L====
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[[Image:Chiba_talks_JW1908_30_RS1_01.gif|thumb|left|'''Fig.5'''  <br>E.coli strain,Senders:JW1908,BBa_T9002:JW1908,30°C,Receiver cells/Sender cells = 1.]]
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[[Image:Chiba_talks_JW1908_30_RS1_01.gif|thumb|left|'''Fig.5'''  <br>E.coli strain,Senders:JW1908,BBa_T9002:JW1908,30&deg;C.All measurements are averages from three replicate cultures with error bars representing standard deviations.]]
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[[Image:Chiba_talks_JW1908_30_RS1_02.gif|thumb|left|'''Fig.6'''  <br>E.coli strain,Senders:JW1908,BBa_T9002:JW1908,30°C,Receiver cells/Sender cells = 1.]]
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[[Image:Chiba_talks_JW1908_30_RS1_02.gif|thumb|left|'''Fig.6'''  <br>E.coli strain,Senders:JW1908,BBa_T9002:JW1908,30&deg;C.All measurements are averages from three replicate cultures with error bars representing standard deviations.]]
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Left:
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*Fig. 6
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#
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*Results
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Right:
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**No significant difference in fluorescence intensity between the cultures containing [http://partsregistry.org/Part:BBa_K084007 BBa_K084007](plac+RhlI) gene transformed cells and the culture containing [http://partsregistry.org/Part:BBa_K084008 BBa_K084008](plac+RhlI(LVA)) gene transformed cells.
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#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.
+
*Discussion
-
#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.
+
**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:100&mu;L,Receiver culture:1000&mu;L====
====Sender culture:100&mu;L,Receiver culture:1000&mu;L====
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[[Image:Chiba_talks_JW1908_30_RS2_01.gif|thumb|left|'''Fig.7'''  <br>E.coli strain,senders:JW1908,BBa_T9002:JW1908,30°C]]
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[[Image:Chiba_talks_JW1908_30_RS2_01.gif|thumb|left|'''Fig.7'''  <br>E.coli strain,senders:JW1908,BBa_T9002:JW1908,30&deg;C.All measurements are averages from three replicate cultures with error bars representing standard deviations.]]
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[[Image:Chiba_talks_JW1908_30_RS2_02.gif|thumb|left|'''Fig.8''' <br>E.coli strain,senders:JW1908,BBa_T9002:JW1908,30°C]]
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[[Image:Chiba_talks_JW1908_30_RS2_02.gif|thumb|left|'''Fig.8''' <br>E.coli strain,senders:JW1908,BBa_T9002:JW1908,30&deg;C.All measurements are averages from three replicate cultures with error bars representing standard deviations.]]
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<br clear=all>
<br clear=all>
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Left:
+
*Fig.7
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#
+
*Results
-
Right:
+
**No significant difference in fluorescence intensity between the culture containing [http://partsregistry.org/Part:BBa_K084007 BBa_K084007](plac+RhlI) gene transformed cells and the culture containing [http://partsregistry.org/Part:BBa_K084008 BBa_K084008](plac+RhlI(LVA)) gene transformed cells.
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#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.
+
*Discussion
-
#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.
+
**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:10&mu;L,Receiver culture:1000&mu;L====
====Sender culture:10&mu;L,Receiver culture:1000&mu;L====
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[[Image:Chiba_talks_JW1908_30_RS3_02.gif|thumb|left|'''Fig.9'''  <br>E.coli strain,senders:JW1908,BBa_T9002:JW1908,30°C,Receiver cells/Sender cells = 100.]]
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[[Image:Chiba_talks_JW1908_30_RS3_02.gif|thumb|left|'''Fig.9'''  <br>E.coli strain,senders:JW1908,BBa_T9002:JW1908,30&deg;C.All measurements are averages from three replicate cultures with error bars representing standard deviations.]]
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*Fig.9
 +
*Results
 +
**The final fluorescence intensity of the culture containing [http://partsregistry.org/Part:BBa_K084008 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.
 +
*Discussion
 +
**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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#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. 
+
<br clear=all>
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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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[[Team:Chiba/Project/Experiments:Sender_Crosstalk|>Back to Sender experiment and result]]
[[Team:Chiba/Project/Experiments:Sender_Crosstalk|>Back to Sender experiment and result]]

Latest revision as of 09:53, 30 October 2008

Chiba-U.gif

Home The Team The Project Parts Submitted to the Registry Reference Notebook Acknowledgements




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.All measurements are averages from three replicate cultures with error bars representing standard deviations.


Sender culture:100μL,Receiver culture:1000μL

Fig.2  
E.coli strain,senders:JW1908,BBa_T9002:JW1908,37°C.All measurements are averages from three replicate cultures with error bars representing standard deviations.



  • Fig. 1,Fig. 2
  • Results
    • 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.
    • 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.All measurements are averages from three replicate cultures with error bars representing standard deviations.
Fig.4  
E.coli strain,senders:JW1908,BBa_T9002:JW1908,37°C.All measurements are averages from three replicate cultures with error bars representing standard deviations.


  • Fig. 3
  • Results
    • Response time and final fluorescence intensity showed no significant difference.
  • Discussion
    • We thought AHL concentration was quickly reached the threshold concentration.The fluorescence intensity increased as gfp maturured.
  • Fig. 4
  • Results
    • No significant difference in fluorescence intensity between the culture containing [http://partsregistry.org/Part:BBa_K084007 BBa_K084007](plac+RhlI) gene transformed cells and the culture containing [http://partsregistry.org/Part:BBa_K084008 BBa_K084008](plac+RhlI(LVA)) gene transformed cells.
  • Discussion
    • 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.All measurements are averages from three replicate cultures with error bars representing standard deviations.
Fig.6  
E.coli strain,Senders:JW1908,BBa_T9002:JW1908,30°C.All measurements are averages from three replicate cultures with error bars representing standard deviations.


  • Fig. 6
  • Results
    • No significant difference in fluorescence intensity between the cultures containing [http://partsregistry.org/Part:BBa_K084007 BBa_K084007](plac+RhlI) gene transformed cells and the culture containing [http://partsregistry.org/Part:BBa_K084008 BBa_K084008](plac+RhlI(LVA)) gene transformed cells.
  • Discussion
    • 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.All measurements are averages from three replicate cultures with error bars representing standard deviations.
Fig.8 
E.coli strain,senders:JW1908,BBa_T9002:JW1908,30°C.All measurements are averages from three replicate cultures with error bars representing standard deviations.


  • Fig.7
  • Results
    • No significant difference in fluorescence intensity between the culture containing [http://partsregistry.org/Part:BBa_K084007 BBa_K084007](plac+RhlI) gene transformed cells and the culture containing [http://partsregistry.org/Part:BBa_K084008 BBa_K084008](plac+RhlI(LVA)) gene transformed cells.
  • Discussion
    • 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.All measurements are averages from three replicate cultures with error bars representing standard deviations.


  • Fig.9
  • Results
    • The final fluorescence intensity of the culture containing [http://partsregistry.org/Part:BBa_K084008 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.
  • Discussion
    • 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.


>Back to Sender experiment and result