August/6 August 2009

From 2009.igem.org

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===To do in lab===
===To do in lab===
1. Transform & Selection
1. Transform & Selection
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*培地作り
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*Medium buildng
**LB
**LB
**LB Amp+
**LB Amp+
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*Transformation
*Transformation
**About 16 kinds of parts
**About 16 kinds of parts
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     We transform thesefollowing parts today.
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     We transform these following 14 parts today.
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       B0015 Plate 1 23L    S03878 Plate2 16M
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       [http://partsregistry.org/Part:BBa_B0015 B0015] Plate 1 23L    [http://partsregistry.org/Part:BBa_S03878 S03878] Plate2 16M
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       C0179 Pate 2 8M    C0070 Plate2  12H
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       [http://partsregistry.org/Part:BBa_C0179 C0179] Pate 2 8M    <del>C0070 Plate2  12H</del>
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      F1610 Plate2 24G   B0034 Plate1 2M
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      [http://partsregistry.org/Part:BBa_F1610 F1610] Plate2 24G     [http://partsregistry.org/Part:BBa_B0034 B0034] Plate1 2M
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       I0462 Plate1 8O   C0078 Plate1 14D
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       [http://partsregistry.org/Part:BBa_I0462 I0462] Plate1 8O     <del>C0078 Plate1 14D</del>
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      C0077 Plate1 14A   I1466 Plate1 23J  
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      <del>C0077 Plate1 14A</del>    [http://partsregistry.org/Part:BBa_I1466 I1466] Plate1 23J  
        
        
2. Breeding
2. Breeding
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===Design genetic circuits===
===Design genetic circuits===
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We take notice of signal molecules. So I put them in order in the chart below.<br/>
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We have thus far focused our attention on signaling molecules used in quorum sensing systems of bacteria. The ones we will be using are as shown below:<br/>
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　                                                                                                                                 *Inducer ⇒（synthesize)  signal moleculer ⇒（synthesize)  Receiver
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　                                                                                                                               *Inducer ⇒（synthesize) Signal molecule ⇒ Receptor
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  * LuxI ⇒　    3OC6HSL　    　⇒ LuxR
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  * LasI ⇒　　AI-1(3OC12HSL) 　⇒　LasR
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  * LuxI ⇒　    3OC6HSL　    　⇒ LuxR → positive regulation
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  * CinI  ⇒  　3OH,C14:1-HSl  　⇒　CinR
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  * LasI ⇒　　AI-1(3OC12HSL) 　⇒　LasR → positive regulation
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  * RhlI ⇒　　AI-1(C4HSL)  　　 ⇒　RhlR  
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  * CinI  ⇒  　3OH,C14:1-HSl  　⇒　CinR → positive regulation
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  * （exception）agrD synthesize AIP with agrB.AIp synthesize agrC . AgrA which is synthesized by agrC activate promoter.
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  * RhlI ⇒　　AI-1(C4HSL)  　　 ⇒　RhlR → positive regulation
 +
  * AgrB synthesizes AIP from AgrD ⇒ AIP diffuses and attaches to AgrC ⇒ AgrC phosphorylates <br/>and activates AgrA which then promotes transcription at P2 and P3 promoters.
   From now on,We have to do what I mention below
   From now on,We have to do what I mention below
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   1.Search other cell-cell comunication system
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   1.Investigate other cell-cell comunication systems (besides those used in quorum-sensing).
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   2.Check systems component completely
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   2.Check to ensure that system components are working properly.
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   3.Create new systems or reinforce cell funcution with additional ideas
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   3.Create new systems or implement additional cell functions.
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   4.Search the effect of cross talk  
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   4.Investigate the possibility and extent of cross talk between various signaling molecules.
   
   
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           written by Tadasi Nakamura
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           reported by Tadasi Nakamura
 +
 
 +
[https://2009.igem.org/Team:Osaka/NOTES back to NOTES]

Latest revision as of 07:34, 12 October 2009

Contents

Morning Meeting

To do in lab

1. Transform & Selection

  • Medium buildng
    • LB
    • LB Amp+
    • LB Kan+
  • Transformation
    • About 16 kinds of parts 
   We transform these following 14 parts today.
      [http://partsregistry.org/Part:BBa_B0015 B0015] Plate 1 23L    [http://partsregistry.org/Part:BBa_S03878 S03878] Plate2 16M
      [http://partsregistry.org/Part:BBa_C0179 C0179] Pate 2 8M     C0070 Plate2  12H
      [http://partsregistry.org/Part:BBa_F1610 F1610] Plate2 24G     [http://partsregistry.org/Part:BBa_B0034 B0034] Plate1 2M
      [http://partsregistry.org/Part:BBa_I0462 I0462] Plate1 8O      C0078 Plate1 14D
      C0077 Plate1 14A    [http://partsregistry.org/Part:BBa_I1466 I1466] Plate1 23J

2. Breeding

3. Refinement

To bring

  • Timer
  • Pen
  • Slipper

Design genetic circuits

We have thus far focused our attention on signaling molecules used in quorum sensing systems of bacteria. The ones we will be using are as shown below:
　 *Inducer ⇒（synthesize) Signal molecule ⇒ Receptor 

* LuxI ⇒　    3OC6HSL　     　⇒ LuxR → positive regulation
* LasI ⇒　　AI-1(3OC12HSL) 　⇒　LasR → positive regulation
* CinI  ⇒  　3OH,C14:1-HSl  　⇒　CinR → positive regulation
* RhlI ⇒　　AI-1(C4HSL)  　　 ⇒　RhlR → positive regulation
* AgrB synthesizes AIP from AgrD ⇒ AIP diffuses and attaches to AgrC ⇒ AgrC phosphorylates 
and activates AgrA which then promotes transcription at P2 and P3 promoters.

 From now on,We have to do what I mention below
 1.Investigate other cell-cell comunication systems (besides those used in quorum-sensing).
 2.Check to ensure that system components are working properly.
 3.Create new systems or implement additional cell functions.
 4.Investigate the possibility and extent of cross talk between various signaling molecules.

          reported by Tadasi Nakamura

back to NOTES

Retrieved from "http://2009.igem.org/August/6_August_2009"