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

From 2008.igem.org

(Difference between revisions)
(→Sender culture：10μL,Receiver culture：1000μL)
(→Results and Discussion)
Line 30: Line 30:
<br clear=all>
<br clear=all>
#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.
#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.
-
<!-- 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 -->
+
 
#Others responded at the same time(4 hours after induction).Final fluorescence intensity differd.
#Others responded at the same time(4 hours after induction).Final fluorescence intensity differd.
-
<!-- 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 -->
+
 
===Reaction temparature:37°C===
===Reaction temparature:37°C===
Line 42: Line 42:
#Response time and final fluorescence intensity showed no significant difference.
#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.
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:
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.  
#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===
===Reaction temparature:30°C===
-
====センダーの培養液：500&mu;L、レシーバの培養液：500&mu;L====
+
====Sender culture：500&mu;L,Receiver culture：500&mu;L====
[[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.]]
[[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.]]
[[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.]]
[[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.]]
Line 60: Line 56:
Right:
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.  
#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：100&mu;L,Receiver culture：1000&mu;L====
====Sender culture：100&mu;L,Receiver culture：1000&mu;L====
Line 67: Line 63:
<br clear=all>
<br clear=all>
-
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.  
#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：10&mu;L,Receiver culture：1000&mu;L====
====Sender culture：10&mu;L,Receiver culture：1000&mu;L====
Line 78: Line 72:
#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.   
#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.   
-
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.
+
 
[[Team:Chiba/Project/Experiments:Sender_Crosstalk|>Back to Sender experiment and result]]
[[Team:Chiba/Project/Experiments:Sender_Crosstalk|>Back to Sender experiment and result]]

Revision as of 03:59, 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,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


  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：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 started to increase.


>Back to Sender experiment and result

Retrieved from "http://2008.igem.org/Team:Chiba/Sender_experiments/Senders(JW1908)_T9002(JW1908)"