Team:Brasil-SP/Notebook/CharacterizationAssemblies

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<h1>Characterization Assemblies</h1>
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<p> Apart from the main genetic circuit we also assembled others for characterization purposes, such as the validation of the promoters and tunning of our threshold setter concentration, the QteE.</p>
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  <p> Apart from the main genetic circuit we also assembled others for characterization purposes, such as the validation of the promoters and tunning of our threshold setter concentration, the QteE.</p>
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<h2>Promoter BBa_K823003</h2>
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  <h2>Promoter BBa_K823003</h2>
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    <p><strong>Question</strong>: Does the constitutive promoter BBa_K823003 work properly?</p>
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  <p><strong>Question</strong>: Does the constitutive promoter BBa_K823003 work properly?</p>
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<b class="map_link" id="map_link_0" title="" href="https://static.igem.org/mediawiki/2014/9/90/KI.pdf">KI</b>
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       <img src="https://static.igem.org/mediawiki/2014/e/ef/Glowing_E_coli.JPG" width="400px" height="200px"/>
       <img src="https://static.igem.org/mediawiki/2014/e/ef/Glowing_E_coli.JPG" width="400px" height="200px"/>
       <p><strong>Results</strong>: After the incubation period of the transformed <em>E. coli</em> a large portion of the colonies were glowing green. So the promoter does work. Moreover, this biobrick works on <em>E. coli</em> despite the fact it was designed for <em>B. subtilis</em>.</p>
       <p><strong>Results</strong>: After the incubation period of the transformed <em>E. coli</em> a large portion of the colonies were glowing green. So the promoter does work. Moreover, this biobrick works on <em>E. coli</em> despite the fact it was designed for <em>B. subtilis</em>.</p>
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  <h2>Promoter BBa_K143015</h2>
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    <p><strong>Question</strong>: How does the transcription caused by this promoter varies with the IPTG induction?</p>
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  <h2>Tunning of the QteE Threshold</h2>
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    <p><strong>Question</strong>:What are the concentration of QteE needed to hamper the LasR induction of the promoter PlasR?</p>
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    <p>This is the most difficult task of our project. Tunning the production of QteE so that we establish the correct threshold for the discretization of the Cystatin C level in serum. To attack this challenge we designed 3 circuits so that we could plot a calibration curve. In this circuits we put the transcription of the LasR and QteE under two differnt promoters, the Pveg (BBa_K823003) and PlasR (BBa_K143015).</p>
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<b class="map_link_c" id="map_link_0_c" title="" href="https://static.igem.org/mediawiki/2014/b/b3/KII.pdf">KII</b>
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<b class="map_link_c" id="map_link_1_c" title="" href="https://static.igem.org/mediawiki/2014/a/a3/CII.pdf">CII</b>
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<b class="map_link_d" id="map_link_0_d" title="" href="https://static.igem.org/mediawiki/2014/7/78/KIII.pdf">KIII</b>
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<b class="map_link_d" id="map_link_1_d" title="" href="https://static.igem.org/mediawiki/2014/a/a3/CII.pdf">CII</b>
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<b class="map_link_d" id="map_link_2_d" title="" href="https://static.igem.org/mediawiki/2014/9/95/KIV.pdf">AIV</b>
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<b class="map_link_e" id="map_link_3_e" title="" href="https://static.igem.org/mediawiki/2014/9/95/KX.pdf">KX</b>
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Latest revision as of 17:03, 13 October 2014

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Characterization Assemblies

Apart from the main genetic circuit we also assembled others for characterization purposes, such as the validation of the promoters and tunning of our threshold setter concentration, the QteE.

Promoter BBa_K823003

Question: Does the constitutive promoter BBa_K823003 work properly?

KI

Results: After the incubation period of the transformed E. coli a large portion of the colonies were glowing green. So the promoter does work. Moreover, this biobrick works on E. coli despite the fact it was designed for B. subtilis.

Promoter BBa_K143015

Question: How does the transcription caused by this promoter varies with the IPTG induction?

KV BIII

Tunning of the QteE Threshold

Question:What are the concentration of QteE needed to hamper the LasR induction of the promoter PlasR?

This is the most difficult task of our project. Tunning the production of QteE so that we establish the correct threshold for the discretization of the Cystatin C level in serum. To attack this challenge we designed 3 circuits so that we could plot a calibration curve. In this circuits we put the transcription of the LasR and QteE under two differnt promoters, the Pveg (BBa_K823003) and PlasR (BBa_K143015).







KII CII KIV KXI






KIII CII AIV KX






KIII CII AIV KX

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