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<h3 style="background-color:#ce6b9a;color:#ffffff; padding:10px;letter-spacing:1px; margin-top:0;">Capture part </h3> | <h3 style="background-color:#ce6b9a;color:#ffffff; padding:10px;letter-spacing:1px; margin-top:0;">Capture part </h3> | ||
<p><strong>The Capture part is to capture the rare earth ions in the environment and to anchor our cells over the silicon net.</strong></p> | <p><strong>The Capture part is to capture the rare earth ions in the environment and to anchor our cells over the silicon net.</strong></p> | ||
− | <h4><strong>①With appropriate primers, PCR was carried out to amplify target gene | + | <h4><strong>①With appropriate primers, PCR was carried out to amplify target gene oprF-LBT1-12 and oprF-Sitag.</strong> </h4> |
<h4 class="col-xs-12"><strong>②Then we constructed two vectors to achieve expression of LBT and Si-tag.</strong> </h4> | <h4 class="col-xs-12"><strong>②Then we constructed two vectors to achieve expression of LBT and Si-tag.</strong> </h4> | ||
<img title="demo1" src="https://static.igem.org/mediawiki/2017/7/73/2017_HUST_China_Experiment_image2.png" alt="demo1" class="col-xs-4 col-xs-offset-1" style="padding: 10px 0px;"> | <img title="demo1" src="https://static.igem.org/mediawiki/2017/7/73/2017_HUST_China_Experiment_image2.png" alt="demo1" class="col-xs-4 col-xs-offset-1" style="padding: 10px 0px;"> | ||
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<p class="col-xs-12"> | <p class="col-xs-12"> | ||
− | We cloned the sequence of | + | We cloned the sequence of oprF-LBT and Si-tag into the vector pET28α in order to make the sequence be tagged N-terminally with a 6×His tag. 6×His tag is an affinity tag which allows the tagged recombinant protein to be purified in the process of affinity purification. |
</p> | </p> | ||
<h4 class="col-xs-12"><strong>③Detection</strong> </h4> | <h4 class="col-xs-12"><strong>③Detection</strong> </h4> | ||
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<h3 style="background-color:#ce6b9a;color:#ffffff; padding:10px;letter-spacing:1px; margin-top:0;">Construction of the whole circuit</h3> | <h3 style="background-color:#ce6b9a;color:#ffffff; padding:10px;letter-spacing:1px; margin-top:0;">Construction of the whole circuit</h3> | ||
<p> | <p> | ||
− | <I> PmrA-PmrB-Fe-PmrC-GFP</I> All of our part should be built in one plasmid. Therefore, we use endonuclease and ligase to replace the GFP by | + | <I> PmrA-PmrB-Fe-PmrC-GFP</I> All of our part should be built in one plasmid. Therefore, we use endonuclease and ligase to replace the GFP by oprF-LBT and oprF-Si tag. |
</p> | </p> | ||
<img title="demo1" src="https://static.igem.org/mediawiki/2017/5/58/2017_HUST_China_Experiment_image4.png" alt="experiment_circuit" alt="demo1" class="col-xs-4 col-xs-offset-4" style="padding: 10px 0px;"> | <img title="demo1" src="https://static.igem.org/mediawiki/2017/5/58/2017_HUST_China_Experiment_image4.png" alt="experiment_circuit" alt="demo1" class="col-xs-4 col-xs-offset-4" style="padding: 10px 0px;"> |
Revision as of 16:22, 31 October 2017