Team:Grenoble/Biology/Notebook/June/week 26

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<a href="https://2012.igem.org/Team:Grenoble/Biology/Notebook/June">week 23</a>  
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<h1>June</h1>
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<a href="https://2012.igem.org/Team:Grenoble/Biology/Notebook/June/week_24">week 24</a>  
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<a href="https://2012.igem.org/Team:Grenoble/Biology/Notebook/June">Week 23</a> •
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<a href="https://2012.igem.org/Team:Grenoble/Biology/Notebook/June/week_25">week 25</a>  
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<a href="https://2012.igem.org/Team:Grenoble/Biology/Notebook/June/week_24">Week 24</a> •
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<a href="https://2012.igem.org/Team:Grenoble/Biology/Notebook/June/week_26">week 26</a>
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<a href="https://2012.igem.org/Team:Grenoble/Biology/Notebook/June/week_25">Week 25</a> •
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<a href="https://2012.igem.org/Team:Grenoble/Biology/Notebook/June/week_26">Week 26</a>
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<h1> Week 26: June 25<span class="exposant">th</span> to July 01<span class="exposant">st</span> </h1>
<h1> Week 26: June 25<span class="exposant">th</span> to July 01<span class="exposant">st</span> </h1>
<p>For the detection module, we decided to work on a modified membrane receptor. It consists of two merged E. coli membrane receptors. The extracellular part is the extracellular part of Tap, a dipeptide membrane receptor. The cytoplasmic part is the cytoplasmic part of the EnvZ receptor, which has the ability to activate OmpR (by phosphorilation) which is a transcription factor.
<p>For the detection module, we decided to work on a modified membrane receptor. It consists of two merged E. coli membrane receptors. The extracellular part is the extracellular part of Tap, a dipeptide membrane receptor. The cytoplasmic part is the cytoplasmic part of the EnvZ receptor, which has the ability to activate OmpR (by phosphorilation) which is a transcription factor.
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It will be a proof of concept for a bigger system which is a complete pathogene detection module :
It will be a proof of concept for a bigger system which is a complete pathogene detection module :
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<center><img src="https://static.igem.org/mediawiki/2012/5/54/Receptor_2.jpg" alt="receptor_2"/></center>
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<center><img src="https://static.igem.org/mediawiki/2012/5/54/Receptor_2.jpg" alt="receptor_2"/></center></section>
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<h2>1<span class="exposant">st</span> network : with amplifier 1 (RsmA-rsmY)</h2>
<h2>1<span class="exposant">st</span> network : with amplifier 1 (RsmA-rsmY)</h2>
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<center><img src="https://static.igem.org/mediawiki/2012/3/33/Network_RsmA-rsmY.jpg" alt="amplifier_1"/></center>
 
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<h3>Amplifier 1 : RsmA-rsmY system characterisation: (3 final plasmids)</h3>
<h3>Amplifier 1 : RsmA-rsmY system characterisation: (3 final plasmids)</h3>
<h4>Plasmid Mapping</h4>
<h4>Plasmid Mapping</h4>
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<center><img src="https://static.igem.org/mediawiki/2012/7/77/Capture_d%E2%80%99%C3%A9cran_2012-08-06_%C3%A0_12.09.32.png" alt="pLAC_fha1_eCFP"/></center>
<center><img src="https://static.igem.org/mediawiki/2012/7/77/Capture_d%E2%80%99%C3%A9cran_2012-08-06_%C3%A0_12.09.32.png" alt="pLAC_fha1_eCFP"/></center>
On a pSB3C5 plasmid, we wanted to put the construction: pLAC_RBS_RsmA
On a pSB3C5 plasmid, we wanted to put the construction: pLAC_RBS_RsmA
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<center><img src="" alt="pLAC_rsmY"/></center>
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<center><img src="https://static.igem.org/mediawiki/2012/9/97/Plasm_pLAC_RBS_RsmA.jpg" alt="pLAC_rsmY"/></center>
On a pSB1A3 plasmid, we wanted to put the construction: pLAC_rsmY
On a pSB1A3 plasmid, we wanted to put the construction: pLAC_rsmY
<center><img src="https://static.igem.org/mediawiki/2012/2/22/Capture_d%E2%80%99%C3%A9cran_2012-08-06_%C3%A0_12.09.46.png" alt="pLAC_rsmY"/></center>
<center><img src="https://static.igem.org/mediawiki/2012/2/22/Capture_d%E2%80%99%C3%A9cran_2012-08-06_%C3%A0_12.09.46.png" alt="pLAC_rsmY"/></center>
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rsmY (iGEM Grenoble 2011) => final length ≃ 170bp<br/>
rsmY (iGEM Grenoble 2011) => final length ≃ 170bp<br/>
fha1 (iGEM Grenoble 2011) => final length ≃ 80bp<br/>
fha1 (iGEM Grenoble 2011) => final length ≃ 80bp<br/>
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</section>
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<section>
<h2>2<span class="exposant">nd</span> network : with amplifier 2 (cAMP-CRP)</h2>
<h2>2<span class="exposant">nd</span> network : with amplifier 2 (cAMP-CRP)</h2>
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<center><img src="https://static.igem.org/mediawiki/2012/d/d5/Network_cAMP-CRP.jpg" alt="pLAC_rsmY"/></center>
 
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<h3>Amplifier 2: cAMP-CRP system (first construction): (1 final plasmid)</h3>
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<h4>Plasmid Mapping</h4>
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On the pSB4K5 plasmid, we wanted to put the construction: pAra/Bad_RBS_GFP_RBS_Cya
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<center><img src="https://static.igem.org/mediawiki/2012/8/8f/Capture_d%E2%80%99%C3%A9cran_2012-08-06_%C3%A0_14.46.49.png" alt="pLAC_fha1_eCFP"/></center>
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<h4>Biobricks involved</h4>
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pSB4K5 plasmid => final length ≃ 2400bp
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<h4>New parts</h4>
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pAra/Bad_RBS_GFP (Alon collection) => final length ≃ 1300bp <br/>
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RBS_Cya (E. coli chromosom) => final length ≃ 2600bp
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</section>
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Latest revision as of 14:07, 22 August 2012

iGEM Grenoble 2012

iGEM Grenoble iGEM
iGEM 2012
main page
Project

June

Week 23 • Week 24 • Week 25 • Week 26

Week 26: June 25th to July 01st

For the detection module, we decided to work on a modified membrane receptor. It consists of two merged E. coli membrane receptors. The extracellular part is the extracellular part of Tap, a dipeptide membrane receptor. The cytoplasmic part is the cytoplasmic part of the EnvZ receptor, which has the ability to activate OmpR (by phosphorilation) which is a transcription factor.

receptor


It will be a proof of concept for a bigger system which is a complete pathogene detection module :
receptor_2

1st network : with amplifier 1 (RsmA-rsmY)

Amplifier 1 : RsmA-rsmY system characterisation: (3 final plasmids)

Plasmid Mapping

On a pSB4K5 plasmid, we wanted to put the construction: pLAC_fha1_eCFP
pLAC_fha1_eCFP
On a pSB3C5 plasmid, we wanted to put the construction: pLAC_RBS_RsmA
pLAC_rsmY
On a pSB1A3 plasmid, we wanted to put the construction: pLAC_rsmY
pLAC_rsmY

Biobricks involved

pLAC_RBS (BBa_I13601) => final length ≃ 90bp
pLAC (BBa_I13601) => final length ≃ 90bp
eCFP (BBa_E0422 or BBa_E0022) => final length ≃ 800bp
plasmid pSB4K5 => final length ≃ 2400bp
plasmid pSB3C5 => final length ≃ 2400bp
plasmid pSB1A3 => final length ≃ 2400bp

New parts

RsmA (iGEM Grenoble 2011) => final length ≃ 200bp
rsmY (iGEM Grenoble 2011) => final length ≃ 170bp
fha1 (iGEM Grenoble 2011) => final length ≃ 80bp

2nd network : with amplifier 2 (cAMP-CRP)

Amplifier 2: cAMP-CRP system (first construction): (1 final plasmid)

Plasmid Mapping

On the pSB4K5 plasmid, we wanted to put the construction: pAra/Bad_RBS_GFP_RBS_Cya
pLAC_fha1_eCFP

Biobricks involved

pSB4K5 plasmid => final length ≃ 2400bp

New parts

pAra/Bad_RBS_GFP (Alon collection) => final length ≃ 1300bp
RBS_Cya (E. coli chromosom) => final length ≃ 2600bp

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