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| - | <h3 class="title3">XOR1</h3>
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| - | <div class="accordeon">
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| - | <h2 class="faq" title="Click to open the FAQ"> In vitro</h2>
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| - | <div class="infos">
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| - | <img style="width:340px;" src="https://static.igem.org/mediawiki/2013/2/20/XOR1_petit.jpg" class="imgcontent" />
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| - | <img style="width:340px; float:right;" src="https://static.igem.org/mediawiki/2013/6/6b/XOR1_bio_80.jpg" class="imgcontent" />
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| - | <div class="clear"></div>
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| - | <img style="width:340px; float:left;" src="https://static.igem.org/mediawiki/2013/7/72/Test_carac.jpg" class="imgcontent" />
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| - | <img style="width:340px; float:right;" src="https://static.igem.org/mediawiki/2013/7/78/Graph.jpg" class="imgcontent" />
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| - | <div class="clear"></div>
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| - | <p class="texte"><i>Description des conditions d'expérimentation</i></p>
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| - | <br>
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| - | <p class="texteprotocol"><a href="https://2013.igem.org/Team:INSA_Toulouse/contenu/lab_practice/notebook/protocols/charac_recomb">In vitro recombinase characterization protocol</a></p>
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| - | <img style="width:20px" src="https://static.igem.org/mediawiki/2013/2/23/Top_arrow.png"class="imgcontent2" /><a href="https://2013.igem.org/Team:INSA_Toulouse/contenu/lab_practice/results">Back to the top</a></p>
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| - | <div class="accordeon">
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| - | <h2 class="faq" title="Click to open the FAQ"> In vivo</h2>
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| - | <div class="infos">
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| - | <img style="width:340px; float:left;" src="https://static.igem.org/mediawiki/2013/2/20/XOR1_petit.jpg" class="imgcontent" />
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| - | <img style="width:340px; float:right;" src="https://static.igem.org/mediawiki/2013/6/6b/XOR1_bio_80.jpg" class="imgcontent" />
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| - | <div class="clear"></div>
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| - | <img style="width:340px; float:left;" src="https://static.igem.org/mediawiki/2013/7/72/Test_carac.jpg" class="imgcontent" />
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| - | <img style="width:340px; float:right;" src="https://static.igem.org/mediawiki/2013/7/78/Graph.jpg" class="imgcontent" />
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| - | <div class="clear"></div>
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| - | <p class="texte"><i>Description des conditions d'expérimentation</i></p>
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| - | <br>
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| - | <p class="texteprotocol"><a href="https://2013.igem.org/Team:INSA_Toulouse/contenu/lab_practice/notebook/protocols/charac_recomb">In vitro recombinase characterization protocol</a></p>
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| - | <img style="width:20px" src="https://static.igem.org/mediawiki/2013/2/23/Top_arrow.png"class="imgcontent2" /><a href="https://2013.igem.org/Team:INSA_Toulouse/contenu/lab_practice/results">Back to the top</a></p>
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| - | <h3 class="title3">AND1</h3>
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| - | <h2 class="faq" title="Click to open the FAQ"> In vitro</h2>
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| - | <div class="infos">
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| - | <img style="width:340px; float:left;" src="https://static.igem.org/mediawiki/2013/2/20/XOR1_petit.jpg" class="imgcontent" />
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| - | <img style="width:340px; float:right;" src="https://static.igem.org/mediawiki/2013/6/6b/XOR1_bio_80.jpg" class="imgcontent" />
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| - | <div class="clear"></div>
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| - | <img style="width:340px; float:left;" src="https://static.igem.org/mediawiki/2013/7/72/Test_carac.jpg" class="imgcontent" />
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| - | <img style="width:340px; float:right;" src="https://static.igem.org/mediawiki/2013/7/78/Graph.jpg" class="imgcontent" />
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| - | <div class="clear"></div>
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| - | <p class="texte"><i>Description des conditions d'expérimentation</i></p>
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| - | <br>
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| - | <br>
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| - | <p class="texteprotocol"><a href="https://2013.igem.org/Team:INSA_Toulouse/contenu/lab_practice/notebook/protocols/charac_recomb">In vitro recombinase characterization protocol</a></p>
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| - | <img style="width:20px" src="https://static.igem.org/mediawiki/2013/2/23/Top_arrow.png"class="imgcontent2" /><a href="https://2013.igem.org/Team:INSA_Toulouse/contenu/lab_practice/results">Back to the top</a></p>
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| - | <h2 class="faq" title="Click to open the FAQ"> In vivo</h2>
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| - | <div class="infos">
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| - | <img style="width:340px; float:left;" src="https://static.igem.org/mediawiki/2013/2/20/XOR1_petit.jpg" class="imgcontent" />
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| - | <img style="width:340px; float:right;" src="https://static.igem.org/mediawiki/2013/6/6b/XOR1_bio_80.jpg" class="imgcontent" />
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| - | <div class="clear"></div>
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| - | <img style="width:340px; float:left;" src="https://static.igem.org/mediawiki/2013/7/72/Test_carac.jpg" class="imgcontent" />
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| - | <img style="width:340px; float:right;" src="https://static.igem.org/mediawiki/2013/7/78/Graph.jpg" class="imgcontent" />
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| - | <div class="clear"></div>
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| - | <p class="texte"><i>Description des conditions d'expérimentation</i></p>
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| - | <br>
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| - | <br>
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| - | <p class="texteprotocol"><a href="https://2013.igem.org/Team:INSA_Toulouse/contenu/lab_practice/notebook/protocols/charac_recomb">In vitro recombinase characterization protocol</a></p>
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| - | <img style="width:20px" src="https://static.igem.org/mediawiki/2013/2/23/Top_arrow.png"class="imgcontent2" /><a href="https://2013.igem.org/Team:INSA_Toulouse/contenu/lab_practice/results">Back to the top</a></p>
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| - | <h3 class="title3">Red Light Sensor</h3>
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| - | <div class="accordeon">
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| - | <h2 class="faq" title="Click to open the FAQ"> In vitro</h2>
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| - | <div class="infos">
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| - | <p class="texte"><b>Objective</b><br>
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| - | Characterize the red light sensor system using Cph8 with its promoter pOmpc. Reminder: in darkness, the EnvZ histidine kinase of Cph8 phosphorylates endogenous OmpR, a transcription factor which activates transcription from the OmpC promoter. Under red light conditions, the activity of the EnvZ histidine kinase domain is inhibited, impeaching transcription from the downstream sequence. <br>
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| - | <b>Conception </b><br>
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| - | The following construction was designed:<br>
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| - | <img src="https://static.igem.org/mediawiki/2013/5/52/Red_sensor80.png" class="imgcontent" />
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| - | <p class="texte"> Cph8 gene was under the control of the pTet promoter, the general inducer system to mimic the real behavior of the red light sensor system in the E. calculus design (https://2013.igem.org/Team:INSA_Toulouse). The transformed strain was supposed to express the modified RFP while in the dark. In red light conditions, expression was supposedly promoted, except when aTc was added in the media.<br>
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| - | <b>Result </b><br>
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| - | We obtain the previous construction to characterize the red light sensor system.<br>
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| - | Used parts are available here:<br>
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| - | - <a href="http://parts.igem.org/Part:BBa_K608002">Strong promoter and strong rbs</a><br>
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| - | - <a href="http://parts.igem.org/Part:BBa_I15008">Ho1</a><br>
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| - | - <a href="http://parts.igem.org/Part:BBa_K081017">PcyA</a><br>
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| - | - <a href="http://parts.igem.org/Part:BBa_P0440">TetR</a><br>
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| - | - <a href="http://parts.igem.org/Part:BBa_R0082">pOmpC promoter</a><br>
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| - | - <a href="http://parts.igem.org/Part:BBa_K081014">rfp</a><br>
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| - | - <a href="http://parts.igem.org/Part:BBa_R0040">ptet</a><br>
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| - | - <a href="http://parts.igem.org/Part:BBa_K592018">Cph8</a><br>
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| - | <b>Discussion </b><br>
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| - | We succeeded in cloning the <a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1132017">construction</a>, but did not characterize it due to lack of time. The final biobrick must be used in an E. coli deficient in wild-type EnvZ. The delta EnvZ strain was obtained from the Paris Bettencourt team and has been already described by many other iGEM projects. <br>
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| - | <a href="http://www.nature.com/nature/journal/v438/n7067/abs/nature04405.html">http://www.nature.com/nature/journal/v438/n7067/abs/nature04405.html</a><br>
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| - | </p>
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| - | <img style="width:20px" src="https://static.igem.org/mediawiki/2013/2/23/Top_arrow.png"class="imgcontent2" /><a href="https://2013.igem.org/Team:INSA_Toulouse/contenu/lab_practice/results">Back to the top</a></p>
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| - | <h3 class="title3">Blue Light Sensor</h3>
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| - | <div class="accordeon">
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| - | <h2 class="faq" title="Click to open the FAQ"> In vitro</h2>
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| - | <div class="infos">
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| - | <img style="width:340px; float:left;" src="https://static.igem.org/mediawiki/2013/2/20/XOR1_petit.jpg" class="imgcontent" />
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| - | <img style="width:340px; float:right;" src="https://static.igem.org/mediawiki/2013/6/6b/XOR1_bio_80.jpg" class="imgcontent" />
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| - | <div class="clear"></div>
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| - | <img style="width:340px; float:left;" src="https://static.igem.org/mediawiki/2013/7/72/Test_carac.jpg" class="imgcontent" />
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| - | <img style="width:340px; float:right;" src="https://static.igem.org/mediawiki/2013/7/78/Graph.jpg" class="imgcontent" />
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| - | <div class="clear"></div>
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| - | <p class="texte"><i>Description des conditions d'expérimentation</i></p>
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| - | <br>
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| - | <br>
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| - | <p class="texteprotocol"><a href="https://2013.igem.org/Team:INSA_Toulouse/contenu/lab_practice/notebook/protocols/charac_recomb">In vitro recombinase characterization protocol</a></p>
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| - | <br>
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| - | <img style="width:20px" src="https://static.igem.org/mediawiki/2013/2/23/Top_arrow.png"class="imgcontent2" /><a href="https://2013.igem.org/Team:INSA_Toulouse/contenu/lab_practice/results">Back to the top</a></p>
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| - | </div>
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| - | </div>
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| - | <div class="accordeon">
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| - | <h2 class="faq" title="Click to open the FAQ"> In vivo</h2>
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| - | <div class="infos">
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| - | <img style="width:340px; float:left;" src="https://static.igem.org/mediawiki/2013/2/20/XOR1_petit.jpg" class="imgcontent" />
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| - | <img style="width:340px; float:right;" src="https://static.igem.org/mediawiki/2013/6/6b/XOR1_bio_80.jpg" class="imgcontent" />
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| - | <div class="clear"></div>
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| - | <img style="width:340px; float:left;" src="https://static.igem.org/mediawiki/2013/7/72/Test_carac.jpg" class="imgcontent" />
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| - | <img style="width:340px; float:right;" src="https://static.igem.org/mediawiki/2013/7/78/Graph.jpg" class="imgcontent" />
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| - | <div class="clear"></div>
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| - | <p class="texte"><i>Description des conditions d'expérimentation</i></p>
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| - |
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| - | <br>
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| - | <br>
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| - | <p class="texteprotocol"><a href="https://2013.igem.org/Team:INSA_Toulouse/contenu/lab_practice/notebook/protocols/charac_recomb">In vitro recombinase characterization protocol</a></p>
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| - | <br>
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| - | <img style="width:20px" src="https://static.igem.org/mediawiki/2013/2/23/Top_arrow.png"class="imgcontent2" /><a href="https://2013.igem.org/Team:INSA_Toulouse/contenu/lab_practice/results">Back to the top</a></p>
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| - | <h3 class="title3">Pol T7</h3>
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| - | <div class="accordeon">
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| - | <h2 class="faq" title="Click to open the FAQ"> In vitro</h2>
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| - | <div class="infos">
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| - | <img style="width:340px; float:left;" src="https://static.igem.org/mediawiki/2013/2/20/XOR1_petit.jpg" class="imgcontent" />
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| - | <img style="width:340px; float:right;" src="https://static.igem.org/mediawiki/2013/6/6b/XOR1_bio_80.jpg" class="imgcontent" />
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| - | <div class="clear"></div>
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| - | <img style="width:340px; float:left;" src="https://static.igem.org/mediawiki/2013/7/72/Test_carac.jpg" class="imgcontent" />
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| - | <img style="width:340px; float:right;" src="https://static.igem.org/mediawiki/2013/7/78/Graph.jpg" class="imgcontent" />
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| - | <div class="clear"></div>
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| - | <p class="texte"><i>Description des conditions d'expérimentation</i></p>
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| - |
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| - | <br>
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| - | <br>
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| - | <p class="texteprotocol"><a href="https://2013.igem.org/Team:INSA_Toulouse/contenu/lab_practice/notebook/protocols/charac_recomb">In vitro recombinase characterization protocol</a></p>
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| - | <br>
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| - | <img style="width:20px" src="https://static.igem.org/mediawiki/2013/2/23/Top_arrow.png"class="imgcontent2" /><a href="https://2013.igem.org/Team:INSA_Toulouse/contenu/lab_practice/results">Back to the top</a></p>
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| - | <br>
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| - | </div>
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| - | </div>
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| - |
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| - | <div class="accordeon">
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| - |
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| - | <h2 class="faq" title="Click to open the FAQ"> In vivo</h2>
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| - | <div class="infos">
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| - | <img style="width:340px; float:left;" src="https://static.igem.org/mediawiki/2013/2/20/XOR1_petit.jpg" class="imgcontent" />
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| - | <img style="width:340px; float:right;" src="https://static.igem.org/mediawiki/2013/6/6b/XOR1_bio_80.jpg" class="imgcontent" />
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| - | <div class="clear"></div>
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| - | <img style="width:340px; float:left;" src="https://static.igem.org/mediawiki/2013/7/72/Test_carac.jpg" class="imgcontent" />
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| - | <img style="width:340px; float:right;" src="https://static.igem.org/mediawiki/2013/7/78/Graph.jpg" class="imgcontent" />
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| - | <div class="clear"></div>
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| - | <p class="texte"><i>Description des conditions d'expérimentation</i></p>
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| - |
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| - | <br>
| |
| - | <br>
| |
| - | <p class="texteprotocol"><a href="https://2013.igem.org/Team:INSA_Toulouse/contenu/lab_practice/notebook/protocols/charac_recomb">In vitro recombinase characterization protocol</a></p>
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| - | <br>
| |
| - | <img style="width:20px" src="https://static.igem.org/mediawiki/2013/2/23/Top_arrow.png"class="imgcontent2" /><a href="https://2013.igem.org/Team:INSA_Toulouse/contenu/lab_practice/results">Back to the top</a></p>
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| - | <br>
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| - | <br>
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| - | </div>
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| - | </div>
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| - |
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| - |
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| - |
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| - | <h3 class="title3">General inductor</h3>
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| - |
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| - | <div class="accordeon">
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| - |
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| - | <h2 class="faq" title="Click to open the FAQ"> In vivo</h2>
| |
| - | <div class="infos">
| |
| - |
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| - | <img src="https://static.igem.org/mediawiki/2013/d/d4/General_inducer_notebook_-_680px.png" class="imgcontent" />
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| - |
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| - | <p class="texte">The couple TetR-pTet system construction is a first step to characterize the entire system. A constitutive promoter (BBa_J23116) was assemble to tetR and then assemble to pTet-rbs-RFP-term (Bba_I13521). pTet is a regulator constitutively ON that expresses the mRFP1 protein. TetR is an inducer that binds to pTet promoter and thus, represses expression the downstream system. Supply of tetracycline or its analog aTc (anhydrotetracycline) is known to bind to tetR and invert the operation (inhibits expression of mRFP in this case).<br>
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| - | <br>
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| - | After 18 hours, clones containing this assembly (BBa_J23116-TetR-pTet-RFP) show a leaky basal expression of mRFP. We suppose that the promoter was too weak to express TetR in large quantity. Assembly of a stronger promoter could improve the system and lock the response to an ON/OFF response. <br>
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| - | <br>
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| - | Besides, an experience was done to analyze the effect of the aTc inducer. Result show a visible induction of the red fluorescent protein expression by addition of aTC (60 ng/mL).</p>
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| - |
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| - | <img src="https://static.igem.org/mediawiki/2013/e/e1/Induct_gen_pfaible.png" class="imgcontent" />
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| - |
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| - | <br>
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| - | <br>
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| - | <img style="width:20px" src="https://static.igem.org/mediawiki/2013/2/23/Top_arrow.png"class="imgcontent2" /><a href="https://2013.igem.org/Team:INSA_Toulouse/contenu/lab_practice/results">Back to the top</a></p>
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| - | <br>
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| - | <br>
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| - | </div>
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| - | </div>
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"
