<p>Each team will make new parts during iGEM and will submit them to the Registry of Standard Biological Parts. The iGEM software provides an easy way to present the parts your team has created. The <code><groupparts></code> tag (see below) will generate a table with all of the parts that your team adds to your team sandbox.</p>
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<p>Remember that the goal of proper part documentation is to describe and define a part, so that it can be used without needing to refer to the primary literature. Registry users in future years should be able to read your documentation and be able to use the part successfully. Also, you should provide proper references to acknowledge previous authors and to provide for users who wish to know more.</p>
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<h5>Note</h5>
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<p>Note that parts must be documented on the <a href="http://parts.igem.org/Main_Page"> Registry</a>. This page serves to <i>showcase</i> the parts you have made. Future teams and other users and are much more likely to find parts by looking in the Registry than by looking at your team wiki.</p>
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<h5>Adding parts to the registry</h5>
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<div id="particles-js"></div>
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<p>You can add parts to the Registry at our <a href="http://parts.igem.org/Add_a_Part_to_the_Registry">Add a Part to the Registry</a> link.</p>
<p>We encourage teams to start completing documentation for their parts on the Registry as soon as you have it available. The sooner you put up your parts, the better you will remember all the details about your parts. Remember, you don't need to send us the DNA sample before you create an entry for a part on the Registry. (However, you <b>do</b> need to send us the DNA sample before the Jamboree. If you don't send us a DNA sample of a part, that part will not be eligible for awards and medal criteria.)</p>
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<div class="main" id="parts">
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</div>
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<div class="h1">Parts</div>
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<div class="p">Each part gives us new inspiration.</div>
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<section>
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<table>
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<caption>Part Submissions</caption>
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<tr>
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<div class="column half_size">
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<td>Biobrick</td>
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<td>Part</td>
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<h5>What information do I need to start putting my parts on the Registry?</h5>
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<td>Description</td>
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<p>The information needed to initially create a part on the Registry is:</p>
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</tr>
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<ul>
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<tr>
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<li>Part Name</li>
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<td>BBa_K1981001</td>
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<li>Part type</li>
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<td><i>lsrA</i></td>
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<li>Creator</li>
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<td>Autoinducer-2 import ATP-binding protein LsrA</td>
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<li>Sequence</li>
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</tr>
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<li>Short Description (60 characters on what the DNA does)</li>
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<tr>
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<li>Long Description (Longer description of what the DNA does)</li>
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<td>BBa_K1981002</td>
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<li>Design considerations</li>
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<td><i>lsrB</i></td>
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</ul>
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<td>Autoinducer-2-binding protein LsrB</td>
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</tr>
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<p>
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<tr>
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We encourage you to put up <em>much more</em> information as you gather it over the summer. If you have images, plots, characterization data and other information, please also put it up on the part page. </p>
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<td>BBa_K1981003</td>
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<td><i>lsrC</i></td>
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</div>
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<td>Autoinducer-2 import system permease protein LsrC</td>
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</tr>
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<tr>
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<div class="column half_size">
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<td>BBa_K1981004</td>
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<td><i>lsrD</i></td>
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<h5>Inspiration</h5>
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<td>Autoinducer-2 import system permease protein LsrD</td>
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<p>We have a created a <a href="http://parts.igem.org/Well_Documented_Parts">collection of well documented parts</a> that can help you get started.</p>
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</tr>
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<tr>
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<p> You can also take a look at how other teams have documented their parts in their wiki:</p>
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<td>BBa_K1981005</td>
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<ul>
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<td><i>lsrFG</i></td>
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<li><a href="https://2014.igem.org/Team:MIT/Parts"> 2014 MIT </a></li>
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<td>A protein complex involved in the degradation of phospho-AI-2</td>
<td>A composite part of which GFP expression can directly respond<br> to AI-2 concentration in the natural or artificial environment</td>
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</tr>
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<tr>
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<td>BBa_K1981202</td>
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<td>Autoinducer-2<br> Response Device B</td>
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<td>A composite part which has a tighter regulation and delayed<br> response compared to AI-2 Response Device A</td>
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</tr>
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</table>
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</section>
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<section>
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<div class="h2">1. Best New Basic Part: BBa_K1981101</div>
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<article>
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<div class="h3"><i>lsr</i> promoter of LuxS/AI-2 signaling pathway in <i>E. coli</i> (BBa_K1981101)</div>
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<div class="p">AI-2 is generated by many species of Gram-negative and Gram-positive bacteria. In a group of bacteria exemplified by <i>E. coli</i> MG1655, AI-2 response involves <i>lsr</i> gene clusters that encode <i>lsrACDB</i>, <i>lsrK</i>, <i>lsrFG</i>. <i>plsr</i> is the promoter of the <i>lsr</i> operon.</div>
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<div class="p">We isolated the promoter region of the <i>lsr</i> operon to regulate the expression of the report gene, GFP (BBa_E0040). In order to test the function of the <i>lsr</i> promoter, we transformed the plasmid pTrcHisB containing <i>plsr</i> with GFP gene at its downstream into <i>E. coli</i> MG1655 ΔluxS. We directly add exogenous AI-2 into the culture. The final concentraton of AI-2 is 50μM. Every one hour, optical density was measured and samples were harvested for HPLC analysis. As you can see, after induced by AI-2, the GFP expression is increased compared to control group.</div>
<figcaption>Figure 1.1: GFP expression after promoter <i>lsr</i> is induced by exogenous AI-2</figcaption>
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</figure>
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<div class="p">Then we test whether promoter <i>lsr</i> can respond to different AI-2 concentration. We directly add exogenous AI-2 into the culture. The final concentraton of AI-2 is 50μM, 40μM, 30μM, 20μM, 10μM, 0μM. Every one hour, optical density was measured and samples were harvested for HPLC analysis. The result below shows that promoter <i>lsr</i> can respond to different AI-2 concentration resulting in different GFP expression.</div>
<div class="p">This composite part consists of the AI-2 (autoinducer-2) quorum sensor-inducible promoter BBa_K1981101, a GFP coding sequence BBa_E0040, a double terminator BBa_B0015. We firstly isolated promoter <i>lsr</i> from <i>E.coli</i> MG1655. GFP BBa_E0040 and double terminator BBa_B0015 are standard part offered by iGEM. Then we successfully constrcuted <i>plsr</i>+GFP+double terminator using homologous recombination technology.</div>
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<div class="p">In AI-2 Response Device A, GFP expression is under the control of promoter, <i>lsr</i>. When phospho-AI-2 binds LsrR, expression of GFP ensues. The expression of GFP can directly respond to the AI-2 level in the environment, which is an alternative way to reflect the AI-2 concentration in the natural or artificial environment.</div>
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+
<div class="p">We fisrtly tested whether AI-2 Response Device A can respond to different AI-2 concentration. We directly added exogenous AI-2 into the culture. The final concentraton of AI-2 is 50μM, 40μM, 30μM, 20μM, 10μM, 0μM. Every one hour, optical density was measured and samples were harvested for HPLC analysis. The result below shows that deicve can respond to different AI-2 concentration resulting in different GFP expression.</div>
<figcaption>Figure 2.1: GFP expression of AI-2 Response Device A when add exogenous AI-2.</figcaption>
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</figure>
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<div class="p">AI-2 Consumers was constructed by iGEM 2016 NKU_China by overexpression the components responsible for AI-2 uptake (<i>lsrACDB</i>), phosphorylation (<i>lsrK</i>), and degradation (<i>lsrFG</i>), which can directly absorb and degrade AI-2 in the natural or artificial environment. When E.coli consisting of AI-2 Response Device A are co-cultured with AI-2 Consumers, the GFP expression of AI-2 Response Device A is significantly decreased compared to control group.</div>
<figcaption>Figure 2.2: GFP expression of AI-2 Response Device A when co-cultured with AI-2 Consumers.</figcaption>
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</figure>
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<div class="p">AI-2 Suppliers was constructed by iGEM 2016 NKU_China by overexpressing the components responsible for AI-2 production (<i>luxS</i>, <i>mtn</i>), which can directly supply and enrich the AI-2 molecular level in the natural or artificial environment. When <i>E. coli</i> consisting of AI-2 Response Device A are co-cultured with AI-2 Suppliers, the GFP expression of AI-2 Response Device A is significantly increased compared to control group.</div>
A composite part of which GFP expression can directly respond to AI-2 concentration in the natural or artificial environment
BBa_K1981202
Autoinducer-2 Response Device B
A composite part which has a tighter regulation and delayed response compared to AI-2 Response Device A
1. Best New Basic Part: BBa_K1981101
lsr promoter of LuxS/AI-2 signaling pathway in E. coli (BBa_K1981101)
AI-2 is generated by many species of Gram-negative and Gram-positive bacteria. In a group of bacteria exemplified by E. coli MG1655, AI-2 response involves lsr gene clusters that encode lsrACDB, lsrK, lsrFG. plsr is the promoter of the lsr operon.
We isolated the promoter region of the lsr operon to regulate the expression of the report gene, GFP (BBa_E0040). In order to test the function of the lsr promoter, we transformed the plasmid pTrcHisB containing plsr with GFP gene at its downstream into E. coli MG1655 ΔluxS. We directly add exogenous AI-2 into the culture. The final concentraton of AI-2 is 50μM. Every one hour, optical density was measured and samples were harvested for HPLC analysis. As you can see, after induced by AI-2, the GFP expression is increased compared to control group.
Figure 1.1: GFP expression after promoter lsr is induced by exogenous AI-2
Then we test whether promoter lsr can respond to different AI-2 concentration. We directly add exogenous AI-2 into the culture. The final concentraton of AI-2 is 50μM, 40μM, 30μM, 20μM, 10μM, 0μM. Every one hour, optical density was measured and samples were harvested for HPLC analysis. The result below shows that promoter lsr can respond to different AI-2 concentration resulting in different GFP expression.
Figure 1.2: GFP expression when add exogenous AI-2
2. Best New Composite Part: BBa_K1981201
Autoinducer-2 Response Device A
This composite part consists of the AI-2 (autoinducer-2) quorum sensor-inducible promoter BBa_K1981101, a GFP coding sequence BBa_E0040, a double terminator BBa_B0015. We firstly isolated promoter lsr from E.coli MG1655. GFP BBa_E0040 and double terminator BBa_B0015 are standard part offered by iGEM. Then we successfully constrcuted plsr+GFP+double terminator using homologous recombination technology.
In AI-2 Response Device A, GFP expression is under the control of promoter, lsr. When phospho-AI-2 binds LsrR, expression of GFP ensues. The expression of GFP can directly respond to the AI-2 level in the environment, which is an alternative way to reflect the AI-2 concentration in the natural or artificial environment.
We fisrtly tested whether AI-2 Response Device A can respond to different AI-2 concentration. We directly added exogenous AI-2 into the culture. The final concentraton of AI-2 is 50μM, 40μM, 30μM, 20μM, 10μM, 0μM. Every one hour, optical density was measured and samples were harvested for HPLC analysis. The result below shows that deicve can respond to different AI-2 concentration resulting in different GFP expression.
Figure 2.1: GFP expression of AI-2 Response Device A when add exogenous AI-2.
AI-2 Consumers was constructed by iGEM 2016 NKU_China by overexpression the components responsible for AI-2 uptake (lsrACDB), phosphorylation (lsrK), and degradation (lsrFG), which can directly absorb and degrade AI-2 in the natural or artificial environment. When E.coli consisting of AI-2 Response Device A are co-cultured with AI-2 Consumers, the GFP expression of AI-2 Response Device A is significantly decreased compared to control group.
Figure 2.2: GFP expression of AI-2 Response Device A when co-cultured with AI-2 Consumers.
AI-2 Suppliers was constructed by iGEM 2016 NKU_China by overexpressing the components responsible for AI-2 production (luxS, mtn), which can directly supply and enrich the AI-2 molecular level in the natural or artificial environment. When E. coli consisting of AI-2 Response Device A are co-cultured with AI-2 Suppliers, the GFP expression of AI-2 Response Device A is significantly increased compared to control group.
Figure 2.3: GFP expression of AI-2 Response Device A when co-cultured with AI-2 Suppliers.
