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We used the original <I> <I> PmrA-PmrB-Fe </I> </I>-<I> <I> <I> <I> PmrC-GFP </I></I></I></I>circuit to test whether our | We used the original <I> <I> PmrA-PmrB-Fe </I> </I>-<I> <I> <I> <I> PmrC-GFP </I></I></I></I>circuit to test whether our | ||
PmrABC system works in our plasmid, then the expression and efficacy | PmrABC system works in our plasmid, then the expression and efficacy | ||
− | of LBT1-12 were tested . | + | of <I>LBT1-12</I> were tested . |
</p> | </p> | ||
</div> | </div> | ||
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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 oprF-LBT1-12 and oprF-Sitag.</strong> </h4> | + | <h4><strong>①With appropriate primers, PCR was carried out to amplify target gene <I>oprF-LBT1-12</I> and <I>oprF-Sitag</I>.</strong> </h4> |
− | <h4 class="col-xs-12"><strong>②Then we constructed two vectors to achieve expression of LBT and | + | <h4 class="col-xs-12"><strong>②Then we constructed two vectors to achieve expression of <I>LBT</I> and <I>Sitag</I>.</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;"> | ||
<img title="demo1" src="https://static.igem.org/mediawiki/2017/7/78/2017_HUST_China_Experiment_image3.png" alt="experiment_circuit" alt="demo1" class="col-xs-4 col-xs-offset-2" style="padding: 10px 0px;"> | <img title="demo1" src="https://static.igem.org/mediawiki/2017/7/78/2017_HUST_China_Experiment_image3.png" alt="experiment_circuit" alt="demo1" class="col-xs-4 col-xs-offset-2" style="padding: 10px 0px;"> | ||
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<p class="col-xs-12"> | <p class="col-xs-12"> | ||
− | We cloned the sequence of oprF-LBT and | + | We cloned the sequence of <I>oprF-LBT</I> and <I>Sitag</I> 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> | ||
<p class="col-xs-12"> | <p class="col-xs-12"> | ||
− | In the meanwhile,we added the FLAG tag to achieve that whether the LBTs and the | + | In the meanwhile, we added the FLAG tag to achieve that whether the <I>LBTs</I> and the <I>Sitag</I> was expressed on the cell membrane. |
</p> | </p> | ||
</div> | </div> |
Revision as of 18:44, 31 October 2017