Team:Paris/Construction

From 2008.igem.org

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|align="center"| [[Team:Paris/September 04|September 04]]
|align="center"| [[Team:Paris/September 04|September 04]]
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| style="background: #D4E2EF;" | FliA
+
| style="background: #D4E2EF;" | [[Image:Icon_coding.png]]<br>FliA
|align=| FliA is a sigma factor involved into the regulation of very various genes, the promoter used in the FIFO were selected to be higly responding to FliA. In order to have the FliA gene possibly used as biobrick, we mutated 2 sites (EcoRI and PstI) inside the FliA sequence by assemby PCR.  
|align=| FliA is a sigma factor involved into the regulation of very various genes, the promoter used in the FIFO were selected to be higly responding to FliA. In order to have the FliA gene possibly used as biobrick, we mutated 2 sites (EcoRI and PstI) inside the FliA sequence by assemby PCR.  
|align="center"| [[Team:Paris/September 04|September 04]]
|align="center"| [[Team:Paris/September 04|September 04]]
|- style="background: #dddddd;"   
|- style="background: #dddddd;"   
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| style="background: #D4E2EF;" | FlhDC
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| style="background: #D4E2EF;" |[[Image:Icon_coding.png]]<br>FlhDC
|align=| FlhDC is the main regulator of the flagella genetic system in E. coli. FlhDC regulates all the promoter of the FIFO.
|align=| FlhDC is the main regulator of the flagella genetic system in E. coli. FlhDC regulates all the promoter of the FIFO.
|align="center"| [[Team:Paris/August 13|August 13]]
|align="center"| [[Team:Paris/August 13|August 13]]
|}
|}

Revision as of 03:43, 30 October 2008

FIFO
Construction Description Validation Date
Icon regulatory.pngIcon rbs.pngIcon rbs.pngIcon reporter.pngPart icon terminator.pngPart icon terminator.png
pFliL(RBS WT)-ECFP tripart (LVA+)
Natural Promoter of E. coli regulated by FlhDC and FliA. The natural rbs of the downstream gene is conserved for a more natural expression of the following gene. The fluorescent marker associated with pFliL is ECFP fused with a LVA tag. October 24
Icon regulatory.pngIcon rbs.pngIcon reporter.png
pFliL(+ RBS WT)-GFP
Natural Promoter of E. coli regulated by FlhDC and FliA. The natural rbs of the downstream gene is conserved for a more natural expression of the following gene. The fluorescent marker associated with pFliL is GFP. October 24
Icon regulatory.pngIcon rbs.pngIcon reporter.pngPart icon terminator.pngPart icon terminator.png
pFlgA(+ RBS WT) - YFP Tripart (LVA+)
Natural Promoter of E. coli regulated by FlhDC and FliA. The natural rbs of the downstream gene is conserved for a more natural expression of the following gene. The fluorescent marker associated with pFlgA is EYFP fused with a LVA tag. August 22
Icon regulatory.pngIcon rbs.pngIcon reporter.pngPart icon terminator.pngPart icon terminator.png
pFlgA(+ RBS WT) - YFP Tripart (LVA-)
Natural Promoter of E. coli regulated by FlhDC and FliA. The natural rbs of the downstream gene is conserved for a more natural expression of the following gene. The fluorescent marker associated with pFlgA is EYFP fused without a LVA tag. August 22
Icon regulatory.pngIcon rbs.pngIcon rbs.pngIcon reporter.pngPart icon terminator.pngPart icon terminator.png
pFlgA(+ RBS WT)-YFP tripart (AAV+)
Natural Promoter of E. coli regulated by FlhDC and FliA. The natural rbs of the downstream gene is conserved for a more natural expression of the following gene. The fluorescent marker associated with pFlgA is EYFP fused with a AAV tag. October 24
Icon regulatory.pngIcon rbs.pngIcon reporter.png
pFlgA(+ RBS WT)-GFP
Natural Promoter of E. coli regulated by FlhDC and FliA. The natural rbs of the downstream gene is conserved for a more natural expression of the following gene. The fluorescent marker associated with pFlgA is GFP. October 24
Icon regulatory.pngIcon rbs.pngIcon rbs.pngIcon reporter.pngPart icon terminator.pngPart icon terminator.png
pFlhB(+ RBS WT)-mRFP tripart (+LVA)
Natural Promoter of E. coli regulated by FlhDC and FliA. The natural rbs of the downstream gene is conserved for a more natural expression of the following gene. The fluorescent marker associated with pFlhB is mRFP with a LVA tag. October 24
Icon regulatory.pngIcon rbs.pngIcon reporter.png
pFlhB(+ RBS WT)-GFP
Natural Promoter of E. coli regulated by FlhDC and FliA. The natural rbs of the downstream gene is conserved for a more natural expression of the following gene. The fluorescent marker associated with pFlhB is GFP. October 26
OSCILLATION
Icon regulatory.pngIcon rbs.pngIcon coding.png
pFliL(+ RBS WT)-EnvZ*
pFliL is regulated by FlhDC and EnvZ regulates pFlhDC thanks to a negative feedback loop, origin of an oscillation behavior. October 26
Icon regulatory.pngPart icon rbs.pngIcon coding.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.pngPart icon terminator.png
pFliL(+ RBS WT)-EnvZ*-GFP generator
The same construction as before but with a GFP in order to follow the dynamic of expression of pFliL. October 29
Icon regulatory.pngIcon rbs.pngIcon coding.png
pFlhB(+ RBS WT)-EnvZ*
pFlhB is regulated by FlhDC and EnvZ regulates pFlhDC thanks to a negative feedback loop, origin of an oscillation behavior. October 24
Icon regulatory.pngPart icon rbs.pngIcon coding.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.pngPart icon terminator.png
pFlhB(+ RBS WT)-EnvZ*-GFP generator
The same construction as before but with a GFP in order to follow the dynamic of expression of pFlhB. October 29
Icon regulatory.pngIcon coding.png
araC/pBad-EnvZ*
A construction made for characterizing the repression power of the mutated EnvZ on our system October 27
SYNCHRONISATION
Part icon regulatory.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.pngPart icon terminator.png
pLas - gfp Tripart
Construction for characterization of the pLas promoter August 1
Part icon regulatory.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.pngPart icon terminator.png
pLas - ECFP
The same as before but with another fluorophore August 1
Icon regulatory.pngIcon rbs.pngIcon coding.pngPart icon terminator.pngPart icon terminator.png
strongest promoter-rbs-lasR-dble ter
A construction for a strong constitutive expression of the Las Regulator in order to get maximal expression in presence of AHL August 20
Icon regulatory.pngIcon rbs.pngIcon coding.pngPart icon terminator.pngPart icon terminator.png
Strong promoter-rbs-lasR-dble ter
A derived construction of the previous one for smaller constitutive expression of LasR in the Bacteria. August 20
Part icon rbs.pngIcon coding.png
Strongest RBS (1)- LasR activator (+LVA)
Intermediary construction. We uses different rbs for Las R in order to get various response dynamic to the presence of AHL August 13
Icon rbs.pngIcon coding.pngPart icon terminator.pngPart icon terminator.png
rbs-lasR + dbl ter
Intermediary construction. We uses different rbs for Las R in order to get various response dynamic to the presence of AHL August 19
Icon coding.pngPart icon terminator.pngPart icon terminator.png
lasR activator + LVA - Double Terminator
Intermediary construction August 1
Part icon rbs.pngIcon coding.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.pngPart icon terminator.png
RBS lasI - GFP tripart
Construction that has to be associated with the FIFO. We will know when LasI is expressed and then when AHL is produced. August 13
Part icon rbs.pngIcon coding.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.pngPart icon terminator.png
RBS-lasI - ECFP
The same as the previous one but with another fluorophore. August 13
NEW GENE AND PROMOTER BIOBRICKS
Part icon regulatory.png
pFliA
Natural promoter of E. coli, extracted by PCR and then inserted in a biobrick vector. pFiA is regulated by FlhDC. October 04
Part icon regulatory.png
pFliL (+RBS WT)
Natural promoter with the wild type rbs of its associated gene in E. coli, extracted by PCR and then inserted in a biobrick vector. pFiA is regulated by FlhDC and FliA. October 10
Part icon regulatory.png
pFlgA (+RBS WT)
Natural promoter with the wild type rbs of its associated gene in E. coli, extracted by PCR and then inserted in a biobrick vector. pFiA is regulated by FlhDC and FliA.. October 10
Part icon regulatory.png
pFlgB (+RBS WT)
Natural promoter with the wild type rbs of its associated gene in E. coli, extracted by PCR and then inserted in a biobrick vector. pFiA is regulated by FlhDC and FliA. October 10
Part icon regulatory.png
pFlhB (+RBS WT)
Natural promoter with the wild type rbs of its associated gene in E. coli, extracted by PCR and then inserted in a biobrick vector. pFiA is regulated by FlhDC and FliA. October 10
Icon coding.png
EnvZ*
EnvZ is a membrane protein involved into the osmotic response and is interacting with OmpR that will bind the promoter of FlhDC ad will repress it. EnvZ* is a mutated form of the coding sequence, giving a phenotype where pFlhDC is completely repressed. EnvZ has been extracted by PVR for the correct mutant strain. September 04
Icon coding.png
FliA
FliA is a sigma factor involved into the regulation of very various genes, the promoter used in the FIFO were selected to be higly responding to FliA. In order to have the FliA gene possibly used as biobrick, we mutated 2 sites (EcoRI and PstI) inside the FliA sequence by assemby PCR. September 04
Icon coding.png
FlhDC
FlhDC is the main regulator of the flagella genetic system in E. coli. FlhDC regulates all the promoter of the FIFO. August 13