August/6 August 2009

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===To do in lab===
===To do in lab===
1. Transform & Selection
1. Transform & Selection
-
*培地作り
+
*Medium buildng
**LB
**LB
**LB Amp+
**LB Amp+
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*Transformation
*Transformation
**About 16 kinds of parts
**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    <del>C0070 Plate2  12H</del>
 +
      [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      <del>C0078 Plate1 14D</del>
 +
      <del>C0077 Plate1 14A</del>    [http://partsregistry.org/Part:BBa_I1466 I1466] Plate1 23J
 +
     
2. Breeding
2. Breeding
*
*
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*Slipper
*Slipper
-
==Design genetic circuits==
+
===Design genetic circuits===
-
We take notice of signal molecules. So I put them in order in the chart below.
+
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/>
 +
　                                                                                                                                *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 <br/>and activates AgrA which then promotes transcription at P2 and P3 promoters.
-
　Indcer  signal moleculer   Receiver
+
   From now on,We have to do what I mention below
-
　　
+
  1.Investigate other cell-cell comunication systems (besides those used in quorum-sensing).
-
  LuxI ⇒　    3OC6HSL　    　⇒ LuxR
+
   2.Check to ensure that system components are working properly.
-
  LasI ⇒　　AI-1(3OC12HSL) 　⇒　LasR
+
  3.Create new systems or implement additional cell functions.
-
   CinI ⇒  　3OH,C14:1-HSl  　⇒　CinR
+
  4.Investigate the possibility and extent of cross talk between various signaling molecules.
 +
   
 +
          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

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