Difference between revisions of "Team:Peking/Description"

 
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            <title>Overview</title>
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        <title></title>
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            <meta name="description" content="Wiki of Peking iGEM 2016" />
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        <meta name="description" content="Wiki of Peking iGEM 2016" />
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            <meta name="author" content="Li Jiamian & Wang Yuqing">
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        <meta name="author" content="Li Jiamian & Wang Yuqing"/>
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         </div>
 
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         <!--/Navigation -->
 
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                 <div class="twelve columns centered text-center">
 
                 <div class="twelve columns centered text-center">
 
                     <h1>Description</h1>
 
                     <h1>Description</h1>
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                     <p class="title1" style="text-align:center">In this section, we describe the background and our Uranium Reaper project in brief. We advise you to have a quick overall  understanding of our project before viewing result pages. </p>
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                     <p class="title1" style="text-align:center">In this section, you could find what we achieved during this summer. For detailed lab results, models or human practices, please use the navigation bar for a quick webpage-redirecting.</p>
 
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                <section>
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                                <div class="texttitle">Overview</div>
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                                 <p class="lead add-bottom" style="color:#5E5656">Uranium (U), which is a radionuclide and heavy metal element, has been released into the environment in increasing amounts, mainly due to activities related to the booming nuclear industry<sup>1</sup>. This has resulted in persistent anthropogenic uranium contamination<sup>2</sup>, which contributes to ecotoxicological problems<sup>3</sup>, environmental degradation<sup>4</sup> and could cause severe health problems. Inhalation, ingestion<sup>5</sup> and skin contact are the main routes of uranium exposure<sup>6</sup>. Once entering human body, uranium tends to accumulate<sup>7</sup> and with its radioactivity and heavy-metal toxicity<sup>8</sup>, often leads to significant adverse health effects on human bodies, including DNA damage<sup>9</sup>, reproductive toxicity<sup>10-12</sup> and nephrotoxicity<sup>13</sup>. Therefore, wastewater containing uranium must be thoroughly treated in order to prevent the release of uranium into the environment<sup>14</sup>.</p>
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                                 <p class="lead add-bottom" style="color:#5E5656">Several physical, biological and chemical methods have been developed for the removal of uranium from aqueous waste. These include physico-chemical methods such as ion-exchange, reverse osmosis, precipitation, flocculation<sup>18</sup>, phytoremediation<sup>1,15</sup>, rhizofiltration and other types of bioremediation<sup>16,17</sup>. However, these methods are often expensive, time-consuming and tedious, or inefficient for the treatment of large volumes of wastewater with low concentrations of the target contaminants<sup>19</sup>. After interviewing the Hunan Nuclear Geology 311 Brigade, a geological exploration unit with Grade A qualification in Hunan Province of China, it became apparent that the favored method of dealing with excavation sites is simply filling the ground with fresh soil and growing appropriate plants on it<sup>20,21</sup>. This minimalist approach is likely to be favored due to the high-cost of alternative methods mentioned above. Hence, efforts are needed to develop suitable alternative technologies to complement or replace the existing methods<sup>19</sup>.</p>
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                                    <h4><a href="javascript:void(0);" onclick="naver('A')">To&nbsp;A</a></h4>
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                                 <p class="lead add-bottom" style="color:#5E5656">To obviate such shortcomings, the Peking iGEM 2016 team developed a novel remediation method, Uranium Reaper, which could remove uranyl ions (the predominant form of aqueous uranium) <sup>15,22,23</sup>, with high efficiency at an affordable cost, thus offering great convenience. Uranium Reaper utilizes a smart covalent crosslinking polymer network which is able to self-assemble in aqueous solution (<a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1989000">BBa_K1989000</a>, <a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1989001">BBa_K1989001</a> and <a href="http://parts.igem.org/Part:BBa_K1989002">BBa_K1989002</a>). The addition of biotin-coated magnetic particles to the solution enables the clearance of the complex self-assembled uranium-containing polymer network by a simple magnet. In this way, uranium pollution is alleviated and the uranyl ions could be cleared and enriched for further use.</p>
+
                                    <h4><a href="javascript:void(0);" onclick="naver('B')">To&nbsp;B</a></h4>
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                                    <h4><a href="javascript:void(0);" onclick="naver('C')">To&nbsp;C</a></h4>
−
                                 <p class="lead add-bottom" style="color:#5E5656">Materials modeled on Uranyl Reaper are not limited to uranium remediation, and could obtain endless functions and applications by attaching different modules of interest to the automatically covalently cross-linking protein network. For example, by replacing SUP with Cadmium-Binding Protein (CBP) or Lead-Binding Protein (LBP) <sup>24,25</sup>, this bio-functional polymer network is capable of adsorbing a variety of heavy metals<sup>26</sup> as confirmed by our experiments. By complementing the leaching and elution circuit in mining, this polymer network would simplify the mining procedures and reduce the amount of contaminated wastewater produced<sup>27,28</sup>. What’s more, by optimizing the number of crosslinking modules, it may be possible to use similar biomaterials for 3D printing. We also aimed to develop a Uranium Reaper Kit, in order to facillitate the use of the material worldwide.</p>
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                                </div>
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                            </div>
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                        </div>
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                                <hr>
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−
                               
+
                       
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                                 <div class="texttitle">General Illustration</div>
+
                       
−
                                  
+
                       
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                                 <figure>
+
                        <div class="nine columns">
−
                                     <p style="text-align:center;"><img style="width:80%;" src="https://static.igem.org/mediawiki/2016/3/36/T--Peking--images_Uranium_Reaper04.png" alt=""/></p>
+
                           
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                                </figure>
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                            <a id="A"></a>
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                                 <hr/>
+
                            <div class="texttitle">Project Achievements</div>
−
                               
+
                            <br/>
−
                                <div class="texttitle">Important sections</div>
+
                            <div class="coll">
−
                                <div class="twelve columns">
+
                                <div class="info">
−
                                 <a href="https://2016.igem.org/Team:Peking/Design"><img src="https://static.igem.org/mediawiki/2016/d/d6/T--Peking--images_description_01.jpg"/></a>
+
                                    <div class="ordi">1.</div>
 +
                                </div>
 +
                                <div class="content">
 +
                                    <p>Constructed a multifunctional protein of interest via molecular biological methods, and introduced the constructed plasmids into the engineered bacteria for protein expression. <a href="https://2016.igem.org/Team:Peking/Basic_Part"/>(Learn more)</a></p>
 +
                                 </div>
 +
                            </div>
 +
                           
 +
                            <div class="coll">
 +
                                 <div class="info">
 +
                                    <div class="ordi">2.</div>
 +
                                </div>
 +
                                <div class="content">
 +
                                    <p>Searched for methods and the best conditions for the extraction of each protein. <a href="https://2016.igem.org/Team:Peking/Notebook/Protocol:purification_of_recombinant_proteins"/>(Learn more)</a> </p>
 +
                                </div>
 +
                            </div>
 +
                           
 +
                            <div class="coll">
 +
                                <div class="info">
 +
                                    <div class="ordi">3.</div>
 +
                                </div>
 +
                                <div class="content">
 +
                                    <p>Demonstrated a quick and stable crosslinking process of Triple SpyTag-SUP and Triple SpyTag-mSA with Triple SpyCatcher via covalent bonds. We also optimized this reaction concerning the relevant parameters such as temperature, pH, etc.. <a href="https://2016.igem.org/Team:Peking/Crosslinking"/>(Learn more)</a> </p>
 +
                                 </div>
 +
                            </div>
 +
                           
 +
                            <div class="coll">
 +
                                 <div class="info">
 +
                                    <div class="ordi">4.</div>
 +
                                </div>
 +
                                <div class="content">
 +
                                    <p>Demonstrated effective adsorption of uranyl ions by monomeric Triple SpyTag-SUP or polymer network containing the SUP module under a number of conditions. The adsorption was highly efficient and fast, not only under experimental conditions but also in simulated seawater or lake water containing uranium pollution. <a href="https://2016.igem.org/Team:Peking/Uranyl-adsorption"/>(Learn more)</a> </p>
 +
                                </div>
 +
                            </div>
 +
                           
 +
                            <div class="coll">
 +
                                <div class="info">
 +
                                    <div class="ordi">5.</div>
 +
                                </div>
 +
                                <div class="content">
 +
                                    <p>Attached biotin to amino-coated magnetic beads and achieved clearance of the polymer network formed via the crosslinking of Triple SpyTag-SUP and Triple SpyTag-mSA with Triple SpyCatcher with a magnet. <a href="https://2016.igem.org/Team:Peking/Clearance"/>(Learn more)</a> </p>
 +
                                </div>
 +
                            </div>
 +
                           
 +
                            <div class="coll">
 +
                                <div class="info">
 +
                                    <div class="ordi">6.</div>
 +
                                </div>
 +
                                <div class="content">
 +
                                    <p>Set up a signal peptide library and screened for optimally suited signal peptides in order to efficiently secrete the proteins of interest. We found two signal peptides of high efficiency - those derived from OmpA and LtIIb. <a href="https://2016.igem.org/Team:Peking/Secretion"/>(Learn more)</a> </p>
 +
                                </div>
 +
                            </div>
 +
                           
 +
                            <div class="coll">
 +
                                <div class="info">
 +
                                    <div class="ordi">7.</div>
 +
                                </div>
 +
                                <div class="content">
 +
                                    <p>Used all the above-mentioned experiments together to demonstrate that the complete Uranium Reaper system, consisting of Triple SpyTag-SUP, Triple SpyTag-mSA, Triple SpyCatcher and biotin-coated magnetic beads, could effectively handle uranium pollution under simulated real-life conditions in about 2 hours. We aimed to optimize this strategy and hoped it could be implemented as a uranyl removal kit. <a href="https://2016.igem.org/Team:Peking/Proof"/>(Learn more)</a> </p>
 +
                                 </div>
 +
                            </div>
 +
                           
 +
                            <div class="coll">
 +
                                 <div class="info">
 +
                                    <div class="ordi">8.</div>
 +
                                </div>
 +
                                <div class="content">
 +
                                    <p>We exchanged the Triple SpyTag-SUP monomer for Triple SpyTag-LBP or Triple SpyTag-CBP, and tried using the same strategy to adsorb lead and cadmium. The results were remarkable, clearly demonstrating that the Uranium Reaper strategy has much potential to be expanded to other heavy metals. <a href="https://2016.igem.org/Team:Peking/Demonstrate"/>(Learn more)</a> </p>
 +
                                 </div>
 +
                            </div>
 +
                            <br/>
 +
                           
 +
                            <div class="coll">
 +
                                 <div class="info">
 +
                                    <div class="ordi">&nbsp;</div>
 +
                                 </div>
 +
                                 <div class="content">
 +
                                     <figure>
 +
                                        <p style="text-align:center;"><img style="width:90%;" src="https://static.igem.org/mediawiki/2016/a/af/T--Peking--image_demotable1.png" alt=""/></p>
 +
                                        <figcaption style="text-align:left;">
 +
                                           
 +
                                        </figcaption>
 +
                                    </figure>
 +
                                 </div>
 +
                            </div>
 +
                           
 +
                            <div class="coll">
 +
                                <div class="info">
 +
                                    <div class="ordi">&nbsp;</div>
 +
                                 </div>
 +
                                <div class="content">
 +
                                    <p>Even though the efficiency of the Uranium Reaper system may be somewhat lower than current methods, it could certainly be optimized through further development work. Importantly, Uranium Reaper is much better in other aspects. In the future, we plan to optimize the entire Uranium Reaper strategy in order to enhance the adsorption efficiency.</p>
 +
                                </div>
 +
                            </div>
  
−
                                 <a href="https://2016.igem.org/Team:Peking/Secretion"><img src="https://static.igem.org/mediawiki/2016/8/8c/T--Peking--images_description_new_02.jpg"/></a>
+
                            <div class="coll">
 +
                                 <div class="info">
 +
                                    <div class="ordi">&nbsp;</div>
 +
                                </div>
 +
                                <div class="content">
 +
                                    <p>For an overall view of our project, please redirect to <a href="https://2016.igem.org/Team:Peking/Description"/>Overview Page</a> or <a href="https://2016.igem.org/Team:Peking/Design"/>Design Page</a>. Links to detailed lab results could also be found on these pages.</p>
 +
                                </div>
 +
                            </div>
  
−
                                <a href="https://2016.igem.org/Team:Peking/Crosslinking"><img src="https://static.igem.org/mediawiki/2016/a/aa/T--Peking--images_description_03.jpg"/></a>
 
  
−
                                <a href="https://2016.igem.org/Team:Peking/Uranyl-adsorption"><img src="https://static.igem.org/mediawiki/2016/5/51/T--Peking--images_description_04.jpg"/></a>
+
                           
−
 
+
                           
−
                                <a href="https://2016.igem.org/Team:Peking/Clearance"><img src="https://static.igem.org/mediawiki/2016/5/55/T--Peking--images_description_new_05.jpg"/></a>
+
                            <p>&nbsp;</p>
−
 
+
                           
−
                                 <a href="https://2016.igem.org/Team:Peking/Demonstrate"><img src="https://static.igem.org/mediawiki/2016/f/f4/T--Peking--images_description_06.jpg"/></a>
+
                           
 +
                           
 +
                           
 +
                           
 +
                            <a id="B"></a>
 +
                           
 +
                            <div class="texttitle">Beyond Experiment</div>
 +
                            <br/>
 +
                            <div class="coll">
 +
                                 <div class="info">
 +
                                    <div class="ordi">1.</div>
 
                                 </div>
 
                                 </div>
−
                               
+
                                 <div class="content">
−
                               
+
                                     <p>We submitted 53 high-quality and well-characterized Standard BioBricks, including a set of derivatives of Triple SpyTag and Triple SpyCatcher, such as the Triple SpyTag-SUP and Triple SpyTag-mSA. <a href="https://2016.igem.org/Team:Peking/Basic_Part"/>(Learn more)</a></p>
−
                               
+
−
                               
+
−
                               
+
−
                                 <div class="references" style="padding-left:24px;padding-right:22px;">
+
−
                                    <h3 >References:</h3>
+
−
                                     <p>
+
−
                                    [1] Piyush Malaviya, Asha Singh. Phytoremediation Strategies for Remediation of Uranium-Contaminated Environments: A Review. Critical Reviews in <i>Environmental Science &amp; Technology</i>, <b>volume 42(24), 2575-2647</b> (2012)
+
−
                                    </p><p>
+
−
                                    [2] Merkel B., Long term fate of uranium tailings in mountain areas. <i>Uranium in the Environment. Springer Berlin Heidelberg</i>, <b>47-56 </b>(2006)
+
−
                                    </p><p>
+
−
                                    [3] Antunes S C, Figueiredo D R D, Marques S M, et al. Evaluation of water column and sediment toxicity from an abandoned uranium mine using a battery of bioassays. <i>Science of the Total Environment</i>, <b>374(2-3):252-259</b> (2007)
+
−
                                    </p><p>
+
−
                                    [4] Boulois H D D, Joner E J, Leyval C, et al. Impact of arbuscular mycorrhizal fungi on uranium accumulation by plants. <i>Journal of Environmental Radioactivity</i>, <b>99(5):775-84</b> (2008)
+
−
                                    </p><p>
+
−
                                    [5] Neves O, Abreu M M, Vicente E M. Uptake of Uranium by Lettuce ( Lactuca sativa L.) in Natural Uranium Contaminated Soils in Order to Assess Chemical Risk for Consumers. <i>Water Air &amp; Soil Pollution</i>, <b>195(1):73-84</b> (2008)
+
−
                                    </p><p>
+
−
                                    [6] Anke M, Seeber O, Müller R, et al. Uranium transfer in the food chain from soil to plants, animals and man. <i>Chemie der Erde - Geochemistry</i>, <b>69(1):75-90</b> (2009)
+
−
                                    </p><p>
+
−
                                    [7] Schnug E, Steckel H, Haneklaus S. Contribution of uranium in drinking waters to the daily uranium intake of humans - A case study from Northern Germany. <i>Landbauforschung Volkenrode</i>, <b>55(4)</b> (2005)
+
−
                                    </p><p>
+
−
                                    [8] Kratz S, Schnug E. Rock phosphates and P fertilizers as sources of U contamination in agricultural soils. <i>Uranium in the Environment</i>. <b>57-67</b> (2006)
+
−
                                    </p><p>
+
−
                                    [9] Chang P, Kim K W, Yoshida S, et al. Uranium accumulation of crop plants enhanced by citric acid. <i>Environmental Geochemistry &amp; Health</i>, <b>27(5-6):529-38</b> (2005)
+
−
                                    </p><p>
+
−
                                    [10] Bednar A J, Medina V F, Ulmer-Scholle D S, et al. Effects of organic matter on the distribution of uranium in soil and plant matrices. <i>Chemosphere</i>, <b>70(2):237-47</b> (2007)
+
−
                                    </p><p>
+
−
                                    [11] Arfsten D P, Still K R, Ritchie G D. A review of the effects of uranium and depleted uranium exposure on reproduction and fetal development. <i>Toxicology &amp; Industrial Health</i>,<b>2001, 17(5-10):180-91</b> (2001)
+
−
                                    </p><p>
+
−
                                    [12] Sztajnkrycer M D, Otten E J. Chemical and radiological toxicity of depleted uranium. <i>Military Medicine</i>, <b>169(3):212-6</b> (2004)
+
−
                                    </p><p>
+
−
                                    [13] Craft E S, Abuqare A W, Flaherty M M, et al. Depleted and natural uranium: chemistry and toxicological effects. <i>Journal of Toxicology &amp; Environmental Health Part B</i>, <b>7(4):297-317</b> (2004)
+
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                                    </p><p>
+
−
                                    [14] Jing B, Yao H, Fan F, et al. Biosorption of uranium by chemically modified Rhodotorula glutinis. <i>Journal of Environmental Radioactivity</i>, <b>101(11):969-973</b> (2010)
+
−
                                    </p><p>
+
−
                                    [15] Stojanovic´, M. D, Stevanovic´, D. R, Milojkovic´, J. V, et al. Phytotoxic Effect of the Uranium on the Growing Up and Development the Plant of Corn. <i>Water Air &amp; Soil Pollution</i>, <b>209(1):401-410</b> (2010)
+
−
                                    </p><p>
+
−
                                    [16] Xie S, Yang J, Chen C, et al. Study on biosorption kinetics and thermodynamics of uranium by Citrobacter freudii. <i>Journal of Environmental Radioactivity</i>, <b>99(1):126-33</b> (2008)
+
−
                                    </p><p>
+
−
                                    [17] Newsome L, Morris K, Lloyd J R. The biogeochemistry and bioremediation of uranium and other priority radionuclides. <i>Chemical Geology</i>, <b>363(1):164-184</b> (2014)
+
−
                                    </p><p>
+
−
                                    [18] Bhalara P D, Punetha D, Balasubramanian K. A review of potential remediation techniques for uranium(VI) ion retrieval from contaminated aqueous environment. <i>Journal of Environmental Chemical Engineering</i>, <b>Journal of Environmental Chemical Engineering</b> (2014)
+
−
                                    </p><p>
+
−
                                    [19] Bhainsa K C, D'Souza S F. Uranium(VI) biosorption by dried roots of Eichhornia crassipes (water hyacinth). <i>Journal of Environmental Science &amp; Health Part A Toxic/hazardous Substances &amp; Environmental Engineering</i>, <b>36(9):1621-31</b> (2001)
+
−
                                    </p><p>
+
−
                                    [20] Neves M O, Figueiredo V R, Abreu M M. Transfer of U, Al and Mn in the water-soil-plant (Solanum tuberosum L.) system near a former uranium mining area (Cunha Baixa, Portugal) and implications to human health. <i>Science of the Total Environment</i>, <b>416(2):156–163</b> (2012)
+
−
                                    </p><p>
+
−
                                    [21] Stojanović M, Pezo L, Lačnjevac Č, et al. Biometric approach in selecting plants for phytoaccumulation of uranium. <i>International Journal of Phytoremediation</i>, <b>18(5):255-262</b> (2015)
+
−
                                    </p><p>
+
−
                                    [22] Kalin M, Wheeler W N, Meinrath G. The removal of uranium from mining waste water using algal/microbial biomass. <i>Journal of Environmental Radioactivity</i>, <b>78(2):151-177</b> (2005)
+
−
                                    </p><p>
+
−
                                    [23] Handley-Sidhu S, Keith-Roach M J, Lloyd J R, et al. A review of the environmental corrosion, fate and bioavailability of munitions grade depleted uranium. <i>Science of the Total Environment</i>, <b>408(23):5690-700</b> (2010)
+
−
                                    </p><p>
+
−
                                    [24] Song L, Caguiat J, Li Z, et al. Engineered single-chain, antiparallel, coiled coil mimics the MerR metal binding site. <i>Journal of Bacteriology</i>, <b>186(6):1861-8</b> (2004)
+
−
                                    </p><p>
+
−
                                    [25] Brown N L, Stoyanov J V, Kidd S P, et al. The MerR family of transcriptional regulators. <i>Fems Microbiology Reviews</i>, <b>27(2-3):145–163</b> (2003)
+
−
                                    </p><p>
+
−
                                    [26] Mulligan C N, Yong R N, Gibbs B F. Remediation technologies for metal-contaminated soils and groundwater: an evaluation. <i>Engineering Geology</i>, <b>60(1-4):193-207 </b>(2001)
+
−
                                    </p><p>
+
−
                                    [27] Campbell K M, Gallegos T J, Landa E R. Biogeochemical aspects of uranium mineralization, mining, milling, and remediation. <i>Applied Geochemistry</i>, <b>57:206-235</b> (2015)
+
−
                                    </p><p>
+
−
                                    [28] Quan, C., Collier, D., Bowell, R.J., van Noort, E.. Optimization of the calcrete-uranium process flowsheet: the implications of metallurgical testwork for the Marenica project. <i>In: Taylor, A. (Ed.), Alta 2010 Uranium Conference, Perth, Australia</i> (2010).
+
−
                                    </p>
+
 
                                 </div>
 
                                 </div>
−
                                  
+
                            </div>
−
                                  
+
                           
−
                             </div><!--twelve row end -->
+
                            <div class="coll">
−
                        </div> <!-- row End -->
+
                                 <div class="info">
−
                    </section> <!-- section end -->
+
                                    <div class="ordi">2.</div>
−
                </div> <!-- primary end -->
+
                                 </div>
−
             </div> <!-- page-content End-->
+
                                <div class="content">
−
         </div> <!-- Content End-->
+
                                    <p>We developed a special software which could be used to calculate the molecular weight distribution of protein polymers using Flory’s theory. The results of testing have demonstrated that the software is accurate and useful. <a href="https://2016.igem.org/Team:Peking/Software"/>(Learn more)</a> </p>
−
       
+
                                </div>
−
       
+
                             </div>
−
       
+
                           
−
       
+
                            <div class="coll">
−
       
+
                                <div class="info">
 +
                                    <div class="ordi">3.</div>
 +
                                </div>
 +
                                <div class="content">
 +
                                    <p>We visited experts from the College of Chemistry and Molecular Engineering and School of Physics of Peking University, respectively, to learn about the current situation surrounding uranium pollution in the real world and how people could control the situation. After finishing the main work, we presented them with the achievements of the project and got their feedback. <a href="https://2016.igem.org/Team:Peking/HP/consulting"/>(Learn more)</a> </p>
 +
                                </div>
 +
                            </div>
 +
                           
 +
                            <div class="coll">
 +
                                <div class="info">
 +
                                    <div class="ordi">4.</div>
 +
                                </div>
 +
                                <div class="content">
 +
                                    <p>We did an interview with the Hunan Nuclear Geology 311 Brigade and gained thorough insights into the treatment of uranyl pollution used by the people on the firing line. This way we could compare the methods they were using with the Uranium Reaper strategy. <a href="https://2016.igem.org/Team:Peking/HP/Gold/311"/>(Learn more)</a> </p>
 +
                                </div>
 +
                            </div>
 +
                           
 +
                            <div class="coll">
 +
                                <div class="info">
 +
                                    <div class="ordi">5.</div>
 +
                                </div>
 +
                                <div class="content">
 +
                                    <p>We helped and collaborated with other iGEM teams by guiding a new team (BHU_China), as well as discussing about project design and technical skills and sharing DNA materials (OUC-China, BIT-China, Tianjin, UCAS, Jinlin_China and BNU-China). <a href="https://2016.igem.org/Team:Peking/Collaborations"/>(Learn more)</a> </p>
 +
                                </div>
 +
                            </div>
 +
                           
 +
                            <div class="coll">
 +
                                <div class="info">
 +
                                    <div class="ordi">6.</div>
 +
                                </div>
 +
                                <div class="content">
 +
                                    <p>We attended the CCiC (Central China iGEM Consortium), which is a large-scale competition-free jamboree of about 50 teams, providing participants with an opportunity for meaningful exchanges of ideas and problem solving. <a href="https://2016.igem.org/Team:Peking/Collaborations"/>(Learn more)</a> </p>
 +
                                </div>
 +
                            </div>
 +
                           
 +
                           
 +
                            <p>&nbsp;</p>
 +
                           
 +
                           
 +
                           
 +
                            <a id="C"></a>
 +
                            <div class="texttitle">Our future plan</div>
 +
                            <br/>
 +
                            <div class="coll">
 +
                                <div class="info">
 +
                                    <div class="ordi">1.</div>
 +
                                </div>
 +
                                <div class="content">
 +
                                    <p>We should reproduce all of the experiments that we have done this summer to make sure the results are credible.</p>
 +
                                </div>
 +
                            </div>
 +
                           
 +
                            <div class="coll">
 +
                                <div class="info">
 +
                                    <div class="ordi">2.</div>
 +
                                </div>
 +
                                <div class="content">
 +
                                    <p>We will optimize the whole strategy to enhance the adsorption efficiency by changing pH, temperature, reaction time of crosslinking and clearance. (The efficiency is only about 60% without further optimization)</p>
 +
                                </div>
 +
                            </div>
 +
                           
 +
                            <div class="coll">
 +
                                <div class="info">
 +
                                    <div class="ordi">3.</div>
 +
                                </div>
 +
                                <div class="content">
 +
                                    <p>According to the results for the adsorption of 13nM uranyl, the polymer network exhibited a good ability in a simulated seawater environment. We could thus also look into other usage scenarios of Uranium Reaper, such as bio-mining and uranium enrichment.</p>
 +
                                </div>
 +
                            </div>
 +
                           
 +
                            <div class="coll">
 +
                                <div class="info">
 +
                                    <div class="ordi">4.</div>
 +
                                </div>
 +
                                <div class="content">
 +
                                    <p>Exchange of the SUP module for other functional proteins. For example, we could integrate proteins which could bind other heavy metals such as mercury so that the polymer network could be used to treat other kinds of pollution as well.</p>
 +
                                </div>
 +
                            </div>
 +
                           
 +
                            <div class="coll">
 +
                                <div class="info">
 +
                                    <div class="ordi">5.</div>
 +
                                </div>
 +
                                <div class="content">
 +
                                    <p>We could assemble enzyme systems behind the SpyTag backbone to create a production plant in vitro. In the protein polymeric network, the concentration of enzymes could be increased and the efficiency of biocatalysis may consequently also be enhanced.</p>
 +
                                </div>
 +
                            </div>
 +
                           
 +
                            <div class="coll">
 +
                                <div class="info">
 +
                                    <div class="ordi">6.</div>
 +
                                </div>
 +
                                <div class="content">
 +
                                    <p>If we optimize the number of SpyTag or SpyCatcher modules per protein monomer, as well as the working concentrations of proteins, we may make protein-3D printing using the Spy Crosslinking Network come true.</p>
 +
                                </div>
 +
                            </div>
 +
                           
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 +
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 +
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Latest revision as of 07:22, 14 October 2018

Description

In this section, you could find what we achieved during this summer. For detailed lab results, models or human practices, please use the navigation bar for a quick webpage-redirecting.

Project Achievements

1.

Constructed a multifunctional protein of interest via molecular biological methods, and introduced the constructed plasmids into the engineered bacteria for protein expression. (Learn more)

2.

Searched for methods and the best conditions for the extraction of each protein. (Learn more)

3.

Demonstrated a quick and stable crosslinking process of Triple SpyTag-SUP and Triple SpyTag-mSA with Triple SpyCatcher via covalent bonds. We also optimized this reaction concerning the relevant parameters such as temperature, pH, etc.. (Learn more)

4.

Demonstrated effective adsorption of uranyl ions by monomeric Triple SpyTag-SUP or polymer network containing the SUP module under a number of conditions. The adsorption was highly efficient and fast, not only under experimental conditions but also in simulated seawater or lake water containing uranium pollution. (Learn more)

5.

Attached biotin to amino-coated magnetic beads and achieved clearance of the polymer network formed via the crosslinking of Triple SpyTag-SUP and Triple SpyTag-mSA with Triple SpyCatcher with a magnet. (Learn more)

6.

Set up a signal peptide library and screened for optimally suited signal peptides in order to efficiently secrete the proteins of interest. We found two signal peptides of high efficiency - those derived from OmpA and LtIIb. (Learn more)

7.

Used all the above-mentioned experiments together to demonstrate that the complete Uranium Reaper system, consisting of Triple SpyTag-SUP, Triple SpyTag-mSA, Triple SpyCatcher and biotin-coated magnetic beads, could effectively handle uranium pollution under simulated real-life conditions in about 2 hours. We aimed to optimize this strategy and hoped it could be implemented as a uranyl removal kit. (Learn more)

8.

We exchanged the Triple SpyTag-SUP monomer for Triple SpyTag-LBP or Triple SpyTag-CBP, and tried using the same strategy to adsorb lead and cadmium. The results were remarkable, clearly demonstrating that the Uranium Reaper strategy has much potential to be expanded to other heavy metals. (Learn more)


 

 

Even though the efficiency of the Uranium Reaper system may be somewhat lower than current methods, it could certainly be optimized through further development work. Importantly, Uranium Reaper is much better in other aspects. In the future, we plan to optimize the entire Uranium Reaper strategy in order to enhance the adsorption efficiency.

 

For an overall view of our project, please redirect to Overview Page or Design Page. Links to detailed lab results could also be found on these pages.

 

Beyond Experiment

1.

We submitted 53 high-quality and well-characterized Standard BioBricks, including a set of derivatives of Triple SpyTag and Triple SpyCatcher, such as the Triple SpyTag-SUP and Triple SpyTag-mSA. (Learn more)

2.

We developed a special software which could be used to calculate the molecular weight distribution of protein polymers using Flory’s theory. The results of testing have demonstrated that the software is accurate and useful. (Learn more)

3.

We visited experts from the College of Chemistry and Molecular Engineering and School of Physics of Peking University, respectively, to learn about the current situation surrounding uranium pollution in the real world and how people could control the situation. After finishing the main work, we presented them with the achievements of the project and got their feedback. (Learn more)

4.

We did an interview with the Hunan Nuclear Geology 311 Brigade and gained thorough insights into the treatment of uranyl pollution used by the people on the firing line. This way we could compare the methods they were using with the Uranium Reaper strategy. (Learn more)

5.

We helped and collaborated with other iGEM teams by guiding a new team (BHU_China), as well as discussing about project design and technical skills and sharing DNA materials (OUC-China, BIT-China, Tianjin, UCAS, Jinlin_China and BNU-China). (Learn more)

6.

We attended the CCiC (Central China iGEM Consortium), which is a large-scale competition-free jamboree of about 50 teams, providing participants with an opportunity for meaningful exchanges of ideas and problem solving. (Learn more)

 

Our future plan

1.

We should reproduce all of the experiments that we have done this summer to make sure the results are credible.

2.

We will optimize the whole strategy to enhance the adsorption efficiency by changing pH, temperature, reaction time of crosslinking and clearance. (The efficiency is only about 60% without further optimization)

3.

According to the results for the adsorption of 13nM uranyl, the polymer network exhibited a good ability in a simulated seawater environment. We could thus also look into other usage scenarios of Uranium Reaper, such as bio-mining and uranium enrichment.

4.

Exchange of the SUP module for other functional proteins. For example, we could integrate proteins which could bind other heavy metals such as mercury so that the polymer network could be used to treat other kinds of pollution as well.

5.

We could assemble enzyme systems behind the SpyTag backbone to create a production plant in vitro. In the protein polymeric network, the concentration of enzymes could be increased and the efficiency of biocatalysis may consequently also be enhanced.

6.

If we optimize the number of SpyTag or SpyCatcher modules per protein monomer, as well as the working concentrations of proteins, we may make protein-3D printing using the Spy Crosslinking Network come true.