Team:Bordeaux/Biobricks

From 2012.igem.org

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<li><a href="http://2012.igem.org/Team:Bordeaux/Introduction">Introduction</a></li>
<li><a href="http://2012.igem.org/Team:Bordeaux/Introduction">Introduction</a></li>
<li><a href="http://2012.igem.org/Team:Bordeaux/Material" >Material & Methods</a></li>
<li><a href="http://2012.igem.org/Team:Bordeaux/Material" >Material & Methods</a></li>
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<li><a href="http://2012.igem.org/Team:Bordeaux/Biobricks">BioBricks</a></li>
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<li><a href="http://2012.igem.org/Team:Bordeaux/Biobricks" class="current">BioBricks</a></li>
<li><a href="http://2012.igem.org/Team:Bordeaux/Construction">Construction</a></li>
<li><a href="http://2012.igem.org/Team:Bordeaux/Construction">Construction</a></li>
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<li><a href="http://2012.igem.org/Team:Bordeaux/Notebook" class="current">Note Book</a></li>
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<li><a href="http://2012.igem.org/Team:Bordeaux/Notebook">Note Book</a></li>
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Revision as of 07:46, 26 September 2012

Note Book - iGEM Bordeaux 2012

iGEM - Bordeaux - Biobricks

RBS-LsrR: BBa_K848000


We added a strong RBS (BBa_B0034) to the LsrR repressor (BBa_K091001) in order to make it a transcriptional unit ready to use. The protein is a repressor of the pLsrA promoter; pAI-2 signal molecule binds to LsrR and inhibits its operation, therefore promoting transcription. LsrK protein (BBa_K848001), which phosphorylates AI-2 signal molecule, is needed for such an inhibition.

RBS-LsrK: BBa_K848001


We added a strong RBS (BBa_B0034) to the LsrK protein (BBa_K091002) in order to make it a transcriptional unit ready to use. The LsrK protein phosphorylates AI-2 signal molecule. Phosphorylated AI-2 will prevent LsrR protein (BBa_K848000) from inhibiting pLsrA promoter.

luxI-lacI-mCherry-dbl terminator : BBa_K848002

We added a double terminator to the intermediate part BBa_S03816 in order to make it a transcriptional unit ready to use. This part Produce LuxI and LacI signal molecules with a mCherry reporter. LuxI is a quorum sensing protein that converts SAM into acyl-homoserine lactone (AHL) thus acting as pLuxR promoter activator when bound to LuxR protein (BBa_K329007). LacI binds to the pLac regulator (BBa_R0010) and PLac01 hybrid regulator (BBa_R0011) inhibits transcription. IPTG binds to LacI and inhibits its operation, therefore promoting transcription.

luxI-lacI-mCherry under lasB /tetO hybrid promoter : BBa_K848003


lasB promoter region is activated by LasR and PAI. The TetO promoter is constitutively ON and repressed by TetR. TetR repression is stopped by the addition of tetracycline. This part produces LuxI and LacI signal molecules with a mCherry reporter. LuxI is a quorum sensing protein that converts SAM into acyl-homoserine lactone (AHL) thus acting as a pLuxR promoter activator when bound to LuxR protein (BBa_K329007)
LacI binds to the pLac regulator (BBa_R0010) and PLlac01 hybrid regulator (BBa_R0011) and inhibits transcription. IPTG binds to LacI and inhibits its operation, therefore promoting transcription.

luxI-lacI-mCherry under LsrA/cI/tetO hybrid promoter : BBa_K848003


The LsrA promoter is repressed by LsrR protein (BBa_K848000). Repression is inhibited by the presence of AI-2 signal molecule phosphorilated by LsrK protein (BBa_K848001) . The cI promoter is repressed by the cI repressor (BBa_C0051)
The TetO promoter is repressed by TetR. TetR repression is inhibited by the addition of tetracycline.
This part produces LuxI and LacI signal molecules with a mCherry reporter. LuxI is a quorum sensing protein that converts SAM into acyl-homoserine lactone (AHL) thus acting a pLuxR promoter activator when bound to LuxR protein (BBa_K329007)
LacI binds to the pLac regulator (BBa_R0010) and PLlac01 hybrid regulator (BBa_R0011) and inhibits transcription. IPTG binds to LacI and inhibits its operation, therefore promoting transcription.