Team:Chiba/Receiver experiments

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(New page: ===LuxR/Plux mutants=== ===AiiA Receiver=== '''Fig.6''' AiiA Receiver '''Design''' オートインデューサー不活性化酵素である...)
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===LuxR/Plux mutants===
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<html><link rel="stylesheet" href="https://2008.igem.org/wiki/index.php?title=User:Hiroki/style.css&action=raw&ctype=text/css" type="text/css" /></html>
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===AiiA Receiver===
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[[Image:Chiba-U.gif]]
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[[Image:AiiA-Receiver Chiba.gif|frame|right|'''Fig.6''' AiiA Receiver]]
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{| style="color:white;background-color:Maroon" cellpadding="3" cellspacing="3" border="1" bordercolor="white" width="100%" align="center"
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!align="center"|[[Team:Chiba|Home]]
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!align="center"|[[Team:Chiba/Team|The Team]]
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!align="center"|[[Team:Chiba/Project|The Project]]
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!align="center"|[[Team:Chiba/Parts|Parts Submitted to the Registry]]
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!align="center"|[[Team:Chiba/Reference|Reference]]
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!align="center"|[[Team:Chiba/Notebook|Notebook]]
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!align="center"|[[Team:Chiba/Acknowledgements|Acknowledgements]]
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|}
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==Receiver crosstalk==
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'''Design'''
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===Design===
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[[Image:Table R chiba.gif|frame|left|Table.1 LuxR family gene]]
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センダーとレシーバーを元来とは異なる組み合わせにすることによって、thresholdに達する時間を遅くする。（杉山）
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オートインデューサー不活性化酵素であるAiiAをレシーバー菌で発現させることにより、レシーバー菌内でのAHL濃度の増加が遅くなるため、AiiAを発現しないレシーバー菌に比べて、GFP発現が遅くなる。
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===Method===
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'''Experiment'''
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===Result===
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*'''Sender'''
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**[http://partsregistry.org/Part:BBa_S03623 BBa_S03623 (AHL auto inucer)]
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[[Image:LuxI-sender Chiba.gif]]
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==Change Receiver's Copy Number==
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===method===
 
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===result===
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===Design===
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*'''Receivers'''
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レシーバーのコピーナンバーを減らすことで、thresholdに達するまでの時間を伸ばす。
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**AiiA Receiver
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（小林）
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[[Image:AiiA-Receiver-genetic-circu Chiba.gif]]
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**non-AiiA Receiver
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===Method===
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[[Image:Non-AiiA-Receiver Chiba.gif]]
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*High Copy Receiver[http://partsregistry.org/Part:BBa_T9002 BBa_T9002 (AHL Reporter)]
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[[Image:T9002.copy-number-change Chiba.gif]]
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*Medium Copy Receiver
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[[Image:Low-copy-Receiver Chiba.gif]]
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High Copy ReceiverとMedium Copy Receiverのディレイタイムはfollowing procedureによってanalyzeした。
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#Transformed sender(Ptet-LuxI), high copy receiver(Ptet-LuxR-Plux-GFP-colE1) and Medium copy receiver(Ptet-LuxR-Plux-GFP-p15A) respectively into E coli strains(BD⊿FliC).
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#Inoculated them independently in liquid media. Incubated at 37℃ 12h.
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#Inoculated again in Fresh liquid media upto about OD600=2 at 37℃
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#Washed sender and receivers.
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#Mixed them. (Sender:Receiver=1000μL:1000μL)
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#Incubated at 30°C.
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#Measured intensity of green fluorescence at regular time intervals.(Fluoroskan AscentR FL＆Fluoroskan AscentR　Thermo ELECTRON CORPORATION)
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===Result===
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'''>[[Team:Chiba/Project#Receiver|Back to the project page]]'''
{| style="color:white;background-color:Maroon" cellpadding="3" cellspacing="3" border="1" bordercolor="white" width="100%" align="center"
{| style="color:white;background-color:Maroon" cellpadding="3" cellspacing="3" border="1" bordercolor="white" width="100%" align="center"

Latest revision as of 19:28, 29 October 2008

Chiba-U.gif

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

Contents

Receiver crosstalk

Design

Table.1 LuxR family gene

センダーとレシーバーを元来とは異なる組み合わせにすることによって、thresholdに達する時間を遅くする。（杉山）

Method

Result

Change Receiver's Copy Number

Design

レシーバーのコピーナンバーを減らすことで、thresholdに達するまでの時間を伸ばす。 （小林）

Method

  • High Copy Receiver[http://partsregistry.org/Part:BBa_T9002 BBa_T9002 (AHL Reporter)]

T9002.copy-number-change Chiba.gif


  • Medium Copy Receiver

Low-copy-Receiver Chiba.gif


High Copy ReceiverとMedium Copy Receiverのディレイタイムはfollowing procedureによってanalyzeした。

  1. Transformed sender(Ptet-LuxI), high copy receiver(Ptet-LuxR-Plux-GFP-colE1) and Medium copy receiver(Ptet-LuxR-Plux-GFP-p15A) respectively into E coli strains(BD⊿FliC).
  2. Inoculated them independently in liquid media. Incubated at 37℃ 12h.
  3. Inoculated again in Fresh liquid media upto about OD600=2 at 37℃
  4. Washed sender and receivers.
  5. Mixed them. (Sender:Receiver=1000μL:1000μL)
  6. Incubated at 30°C.
  7. Measured intensity of green fluorescence at regular time intervals.(Fluoroskan AscentR FL＆Fluoroskan AscentR　Thermo ELECTRON CORPORATION)

Result

>Back to the project page

Home The Team The Project Parts Submitted to the Registry Notebook
Retrieved from "http://2008.igem.org/Team:Chiba/Receiver_experiments"