Team:Paris/Addressing testing

From 2009.igem.org

(Difference between revisions)
(→Adressing system)
(→Adressing system)
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{|
{|
|- style="background: #0a3585; text-align: center; color:white; font-weight:bold; "
|- style="background: #0a3585; text-align: center; color:white; font-weight:bold; "
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|width= 350px| Date
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|width= 350px| row 1
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|width=350px| Authors
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|width=350px| row 2
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|width=350px| Article
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|width=350px| row 3
|- style="background: #bebebe; text-align: center;"
|- style="background: #bebebe; text-align: center;"
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|height=40px| []
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|PCR
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|1999
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|PCR
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|Terry J. Beveridge
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|PCR
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|Structures of gram-negative cell walls and their derived membrane vesicles.
+
 
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|[http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=93954 10438737]
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|- style="background: #d8d8d8; text-align: center;"
|- style="background: #d8d8d8; text-align: center;"
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|2003
|2003
|Miller SI & Guina T.
|Miller SI & Guina T.
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|Bacterial vesicle formation as a mechanism of protein transfer to animals.
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|[http://www.ncbi.nlm.nih.gov/pubmed/14531993?ordinalpos=&itool=EntrezSystem2.PEntrez.Pubmed.Pubmed_ResultsPanel.SmartSearch&log$=citationsensor 14531993]
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|- style="background: #bebebe; text-align: center;"   
|- style="background: #bebebe; text-align: center;"   
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|2005
|2005
|Kuehn MJ & Kesty NC.
|Kuehn MJ & Kesty NC.
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|Bacterial outer membrane vesicles and the host-pathogen interaction.
 
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|[http://www.ncbi.nlm.nih.gov/pubmed/16291643?ordinalpos=&itool=EntrezSystem2.PEntrez.Pubmed.Pubmed_ResultsPanel.SmartSearch&log$=citationsensor 16291643]
 
|- style="background: #d8d8d8; text-align: center;"
|- style="background: #d8d8d8; text-align: center;"
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|2006
|2006
|McBroom AJ & Kuehn MJ.
|McBroom AJ & Kuehn MJ.
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|Outer membrane vesicle production by Escherichia coli is independent of membrane instability.
 
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|[http://www.ncbi.nlm.nih.gov/pubmed/16855227?ordinalpos=&itool=EntrezSystem2.PEntrez.Pubmed.Pubmed_ResultsPanel.SmartSearch&log$=citationsensor 16855227]
 
|- style="background: #bebebe; text-align: center;"   
|- style="background: #bebebe; text-align: center;"   
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|2007
|2007
|McBroom AJ & Kuehn MJ.
|McBroom AJ & Kuehn MJ.
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|Release of outer membrane vesicles by Gram-negative bacteria is a novel envelope stress response.
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|[http://www.ncbi.nlm.nih.gov/pubmed/17163978?ordinalpos=1&itool=EntrezSystem2.PEntrez.Pubmed.Pubmed_ResultsPanel.Pubmed_DefaultReportPanel.Pubmed_RVDocSum 17163978]
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|- style="background: #d8d8d8; text-align: center;"
|- style="background: #d8d8d8; text-align: center;"
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|2009
|2009
|Deatherage BL & Cookson BT.
|Deatherage BL & Cookson BT.
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|Biogenesis of bacterial membrane vesicles
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|[http://www.ncbi.nlm.nih.gov/pubmed/19432795?ordinalpos=1&itool=EntrezSystem2.PEntrez.Pubmed.Pubmed_ResultsPanel.Pubmed_DefaultReportPanel.Pubmed_RVDocSum 19432795]
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|- style="background: #bebebe; text-align: center;"   
|- style="background: #bebebe; text-align: center;"   
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|2009
|2009
|Purnick PE & Weiss R.
|Purnick PE & Weiss R.
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|The second wave of synthetic biology: from modules to systems.
 
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|[http://www.ncbi.nlm.nih.gov/pubmed/19461664?ordinalpos=&itool=EntrezSystem2.PEntrez.Pubmed.Pubmed_ResultsPanel.SmartSearch&log$=citationsensor 19461664]
 
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|}
 
From plate: mRFP
From plate: mRFP

Revision as of 11:27, 17 October 2009

iGEM > Paris > WetLab > Addressing

WetLab

Main - Addressing - Production - reception -
Addressing


Adressing system

global constructions :


plasmid = PSB3T5


Time required : A lOT !!!!!



Experiments ran :

row 1 row 2 row 3
PCR PCR PCR


[] 2003 Miller SI & Guina T.


[] 2005 Kuehn MJ & Kesty NC.
[] 2006 McBroom AJ & Kuehn MJ.
[] 2007 McBroom AJ & Kuehn MJ.


[] 2009 Deatherage BL & Cookson BT.


[] 2009 Purnick PE & Weiss R.

From plate: mRFP


PCR :


ClyA Cterm plasmid : ? Oligo : TM : ClyA Nterm plasmid : ? Oligo : TM : mRFP Cterm plasmid : ? Oligo : TM : mRFP Nterm plasmid : ? Oligo : TM :


Verification on gel

ok

Purification on gel

ok

digestion:

ClyA Cterm S/P RFP Nterm X/P PBAD E/S



ligation:

ClyA Cter-RFP Nter vector PSB1A3

PCR colo :

miniprep:

verifivation of the miniprep by digestion

sequencing [[ClyA]]

-ClyA Nterm= biobrick+sequencage+stock glycerol Done

-ClyA Cterm= biobrick+sequencage+stock glycerol Done

[[-RFP-]]

-RFP Nterm= biobrick+sequencage+stock glycerol Done

-RFP Cterm= biobrick+sequencage+stock glycerol Done

[[-ClyA Cter-RFP Nter-]]

-ClyA Cter-RFP Nter= biobrick+stock glycerol Done

[[-RFP Cter-ClyA Nter-]]

-miss ligation between 2 parts (j'ai fait plusieurs ligation le 7ou8/09, mais pas marché le 9/09 apres culture overnight, voir si les autres ont continué)


[[PBad-ClyA Cter-RFP Nter-]]

-failed many time with:

Pbad cut in S/P and ClyA Cter-RFP Nter in X/P

Pbad cut in E/S and ClyA Cter-RFP Nter in E/X


Export system

Production of protein of interest



Production of protein of interest
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