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| | <div id="main" class="row"> | | <div id="main" class="row"> |
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| - | <a href="https://2014.igem.org/Team:USTC-China/project/killswitch" class="active-a">Kill switch</a> | + | <a href="https://2014.igem.org/Team:USTC-China/project/killswitch" class="active-a">Kill Switch</a> |
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| | <div class="large-9 columns" id="content-page"> | | <div class="large-9 columns" id="content-page"> |
| - | <div class="title"><h1>Kill Switch</h1></div> | + | <div class="title"><h1>Kill Switch</h1> |
| | + | <div data-magellan-expedition="fixed"> |
| | + | <dl class="sub-nav"> |
| | + | <dd data-magellan-arrival="background"><a href="#background">Background</a></dd> |
| | + | <dd data-magellan-arrival="principle"><a href="#principle">Principle</a></dd> |
| | + | </dl> |
| | + | </div> |
| | + | </div> |
| | <div class="text"> | | <div class="text"> |
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| - | <h4 id="background">Background</h4> | + | <a name="background"></a> |
| | + | <h4 data-magellan-destination="background">Background</h4> |
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| | <p>To ensure that our bacteria do no harm to both people and environment, we need to put a set of kill-switch system into the bacteria, so that we can control the growth of them by changing the contents of culture medium.</p> | | <p>To ensure that our bacteria do no harm to both people and environment, we need to put a set of kill-switch system into the bacteria, so that we can control the growth of them by changing the contents of culture medium.</p> |
| | <p>In our project, we trigger the kill switch of <i>E.coli</i> by adding <i>IPTG</i> into the culture. <i>IPTG</i> can induce the expression of gene coding <i>Limulus anti-LPS factor</i> (<i>LALF</i>) downstream the lactose operator, so that the growth of <i>E.coli</i> will be inhibited.<p/> | | <p>In our project, we trigger the kill switch of <i>E.coli</i> by adding <i>IPTG</i> into the culture. <i>IPTG</i> can induce the expression of gene coding <i>Limulus anti-LPS factor</i> (<i>LALF</i>) downstream the lactose operator, so that the growth of <i>E.coli</i> will be inhibited.<p/> |
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| - | <h4 id="principle">Principle</h4> | + | <a name="principle"></a> |
| | + | <h4 data-magellan-destination="principle">Principle</h4> |
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| - | <p>Limulus <i>anti-LPS</i> factor (<i>LALF<i/>) binds LPS on anti-gram negative bacterial surface tightly, forming a biofilm. This can prevent nutrient from entering the cell, causing the growth inhibition. Also, the biofilm can prevent plasmid spreading to the environment and gene pollution.</p> | + | <p>Limulus <i>anti-LPS</i> factor (<i>LALF</i>) binds <i>LPS</i> on anti-gram negative bacterial surface tightly, forming a biofilm. This can prevent nutrient from entering the cell, causing the growth inhibition. Also, the biofilm can prevent plasmid spreading to the environment and gene pollution.</p> |
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| | <p>Besides, to make <i>LALF</i> cross the cell membrane and bind to <i>LPS</i>, we still need an efficient protein export signal sequence of <i>E. coli</i> to lead <i>LALF</i> into extracelluar environment.</p> | | <p>Besides, to make <i>LALF</i> cross the cell membrane and bind to <i>LPS</i>, we still need an efficient protein export signal sequence of <i>E. coli</i> to lead <i>LALF</i> into extracelluar environment.</p> |
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| - | <p><img src="https://static.igem.org/mediawiki/2014/d/da/LAFLG_MEch.jpg" class="th"/></p> | + | <p ><div align="center"><img src="https://static.igem.org/mediawiki/2014/d/da/LAFLG_MEch.jpg" class="th"/ ></div></p> |
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| - | <h4 id="results">Results</h4>
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| - | <p>We constructed the plasmid with lactose operator and signal peptide, and transform this plasmid into E.coil. to test if this kill switch do work. We used gradient plate method in our experiment. As we can see in the picture, the concentration of <i>IPTG</i> in the left of the medium is 0.1% while the concentration on the right is zero. We can see colony only on the right of the medium, the kill switch is effective which illustrates that IPTG can induce the expression of signal peptide so as to inhibit the growth of the <i>E.coil</i>. Also, a decrease of OD is observed after we added <i>IPTG</i> to the liquid culture medium. </p>
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| - | <p><img src="https://static.igem.org/mediawiki/2014/8/87/Sdfsdfs.jpg" class="th"/></p>
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| - | <p>However ,we haven’t connect the LALF gene with signal peptide yet due to the time limit. We will construct the whole plasmid and transform it into <i>E.coil</i>. to see the sterilization effect of LALF in the future work.<p/>
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| | </div> | | </div> |
| | </div> | | </div> |
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