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− | <h1>Our work focuses on three modules: CAAP(Calcium Absorption Peptide) and its secretion, antimicrobial peptide and its improvement and a temperature-dependent kill switch.</h1> | + | <h1 tabIndex="1">It is a story about how we achieve our bone appetit</h1> |
| <div class="column full_size"> | | <div class="column full_size"> |
− | <p>页面摘要,或者是单项奖核心点 Our work focuses on three modules: CAAP(Calcium Absorption Peptide) and its secretion, antimicrobial peptide and its improvement and a temperature-dependent kill switch. </p> | + | <p>This year, we are focusing on the bone health of the elderly in China. To make the elderly pay more attention to their bone health and cooperate with our project, we carried out education through a series of methods to make more elderly people aware of their possible calcium deficiency and know how to protect their bone health. We used synthetic biology to produce engineering bacteria and secrete peptides in the human intestinal tract to promote calcium absorption to improve the calcium absorption rate of the elderly. Simultaneously, we designed the quorum sensing module and improved related parts to ensure that our bacteria can be stable and exist in the human body for a long time and work normally. We also designed and verified the Kill Switch module to ensure safety, created a relevant measurement project to provide a scheme for the elderly to measure their blood calcium concentration. We also explored entrepreneurship to ensure that our project could be used on a larger scale and design the implement so that everyone can use our project easily. We have achieved good results on entrepreneurship so far. </p> |
| </div> | | </div> |
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− | <div class="clear extra_space"></div><!-- 一个章节结束需要clear来留白 如果觉得内容间差异大再加上extra_space --> | + | <div class="clear"></div><!-- 一个章节结束需要clear来留白 如果觉得内容间差异大再加上extra_space --> |
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− | <h2>CAAP expression & secretion</h2>
| + | <h2>Education</h2> |
− | <div class="column two_thirds_size">
| + | <div class="column full_size"> |
− | <img src="https://static.igem.org/mediawiki/2020/8/8b/T--Fudan--map_2.svg" alt="map of calcium intake" />
| + | <p>With the rapid development of science and technology and society, the older generation is gradually falling behind society. Older people face fraud, misinformation, and illness. Lifelong education has become an urgent need for them. However, not everyone can enjoy a good education, so many elderly people in rural areas of China don't even know they are sick. We want the elderly to have a higher quality of life. Although our program can solve many problems, it is not a panacea. We need to educate the elderly to know how to ensure their health. We are committed to exploring more education methods and striving to bring high-quality education to the elderly to prevent diseases and enjoy life.</p> |
− | </div>
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− | <div class="column third_size">
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− | <p>在宽屏,这部分内容占页面1/3;窄屏就是全幅。添加了column同时 图2/3 + 字1/3 正好宽屏的一幅。 We managed to express our designed CAAP(Calcium Absorption Peptide) in E.coli BL21.</p>
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− | </div>
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− | <div class="clear"></div><!-- 一个章节结束需要clear来留白 -->
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− | <h2>每个章节标题用h2 为了搜索引擎全页面只能一个h1标签</h2>
| + | <p>We use previously untapped educational methods to educate older people about biology. Our primary strategy is to educate the younger generation and teach them how to inform their grandparents. Compared with traditional educational strategy, our approach provides emotional support from the younger generation and makes the elderly feel comfortable. To educate young people, in addition to synthetic biology, which we have been teaching for the past few years, we have an online summer camp for high school students to teach elder care. We have developed an online audio course on the basics of biology for all ages. In short, we have made a brave and convincing attempt to carry out scientific communication between the younger generation and a long-neglected social group, the elderly.<p> |
− | <div class="column half_size">
| + | </div> |
− | <img src="https://static.igem.org/mediawiki/2020/8/8b/T--Fudan--map_2.svg" alt="map of calcium intake" />
| + | <div class="clear"></div> |
− | </div>
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− | <div class="column half_size">
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− | <p>在宽屏,这部分内容占页面1/2;窄屏就是全幅。添加了column同时 图1/2 + 字1/2 正好宽屏的一幅。这个代码不在 https://2020.igem.org/Resources/Template_Documentation 但是是支持的 China has an estimated 70 million osteo
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− | porosis patien China has an estimated 70 million osteoporosis patien China has an estimated 70 million osteoporosis patienChina has an estimated 70 million oste
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− | oporosis patienChina has an estimated 70 million osteoporosis patien</p>
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− | </div>
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− | <div class="clear"></div><!-- 一个章节结束需要clear来留白 -->
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− | <h2>每个章节标题用h2 为了搜索引擎全页面只能一个h1标签</h2>
| + | <h2>Kill Switch</h2> |
− | <div class="column third_size">
| + | <div class="column full_size"> |
− | <img src="https://static.igem.org/mediawiki/2020/8/8b/T--Fudan--map_2.svg" alt="map of calcium intake" />
| + | <p>Education ensured that the elderly have an essential awareness of calcium health. We began to experiment, hoping to provide a solution to the bone problems of the elderly in China through synthetic biology. We designed an intestinal microflora that consists of two modules, one for calcium absorption and another for regulating the amount of microflora. But before we do that, we need to consider security first and designed a Kill Switch module. </p> |
− | </div>
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− | <div class="column two_thirds_size">
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− | <p>在宽屏,这部分内容占页面1/3;窄屏就是全幅。添加了column同时 图1/3 + 字2/3 正好宽屏的一幅。 China has an estimated 70 million osteo
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− | porosis patien China has an estimated 70 million osteoporosis patien China has an estimated 70 million osteoporosis patienChina has an estimated 70 million oste
| + | |
− | oporosis patienChina has an estimated 70 million osteoporosis patien</p>
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− | </div>
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− | <div class="clear"></div><!-- 一个章节结束需要clear来留白 -->
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− | <div class="column full_size"><!-- 全幅面的文字内容,上面是否留白可选 -->
| + | <p>The ultimate goal of our project is to produce a commercial calcium supplementary probiotic colonized in the intestines. An eco-friendly commercial probiotic should avoid the occurrence of modified gene vertical transfer and spread widely in the natural environment when excreted from the human intestine. And gene vertical transfer is directly associated with the survival of bacteria. Therefore, we needed to construct a Kill Switch to deprive of the survivability of engineered <i>E. coli</i> outside human bodies. </p> |
− | <p>However, for the Chinese population, the bottlenecks are
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− | as follows:
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− | </p>
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− | <ol>
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− | <li>注意这里列表的写法 页面字会变深 突出内容 ol是编数字的 nts</li>
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− | <li>calcium-supplementing pills daste</li>
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− | <li>The body's absorptioes as calcium intake increases</li>
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− | <li>Vegetables conta列表的写法 页面字会变深 突出内容 ol是编数字的 ul是没有标数字的列表 </li>
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− | </ol>
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− | <p>
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− | However, the currently popular calcium supplement method on the market is either to adjust diet or to consume a lot of calcium supplementing pills. For the elderly, they suffer from poor memory or neglect the importance of calcium pills, often forgetting to consume the supplement; for children, they often refuse to take calcium pills or to maintain some diet high in calcium, thus requiring the "liquid calcium" method to ensure sufficient calcium intake. Ho可以 继续大段内容
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− | </p>
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| + | <p>According to the special situation this year, we have limited time to work in the wet lab. As the Kill Switch model is relatively independent of our main project, we convert to establish the demonstration of the Kill Switch on pure dry lab work. We find out that our Kill Switch design has a very similar basic idea with BNU-China, so we decide to establish collaboration with them. We adopt their raw wet lab experimental data of RelE/RelB system to construct our model and proved our idea. </p> |
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− | <div class="highlight decoration_background decoration_A_top"> | + | <p>We improved the logistic equation, and our model fitted the experimental data from BNU-China well. We performed an efficiency analysis by the adjusted model we had constructed. As our design of BBa_K3606027 employed a more efficient RNA thermometer than that of BNU-China, we changed the properties to simulate the growth curve of our Kill Switch. The result demonstrated that bacteria carrying our Kill Switch were killed more thoroughly in a smaller temperature range, therefore proved that with our Kill Switch, engineered bacteria would have less deleterious effects on the environment (figure 1.). Click <a href="/Team:Fudan/Model">here</a> to see the model of Kill Switch.</p> |
− | <p>
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− | 章节内容的分段就用p开p关,额外需要关注的可以套一个highlight的框,有多种版本
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− | </p>
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| </div> | | </div> |
− | <p>
| + | <div class="row"> |
− | 章节内容的分段就用p开p关
| + | <div class="col m2 l3 hide-on-small-only"> </div> |
− | </p>
| + | <div class="col s12 m8 l6"> |
− | <div class="highlight decoration_B_full"> | + | <img src="https://static.igem.org/mediawiki/2020/2/25/T--Fudan--model2_6.png" alt="model 2_6" style="width:100%"> |
− | <p>
| + | </div> |
− | 章节内容的分段就用p开p关,额外需要关注的可以套一个highlight的框,细看 https://2020.igem.org/Resources/Template_Documentation#highlight | + | |
− | </p> | + | |
| </div> | | </div> |
− | <p>
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− | 章节内容的分段就用p开p关
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− | </p>
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− | <p>
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− | 章节内容的分段就用p开p关 <a href="https://URL"> EXAM 外部链接 href尽量使用https </a>
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− | </p>
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− | <ul>
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− | <li>注意这里列表的写法 页面字会变深 突出内容 ul是没有标数字的列表 nts</li>
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− | <li>calcium-supple menting pills don’t haat taste</li>
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− | <li>Vegetables conta列表的写法 页面字会变深 突出内容 ol是编数字的 ul是没有标数字的列表 </li>
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− | </ul>
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− | <p>
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− | 章节内容的分段就用p开p关
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− | </p>
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− | </div><!-- 每个幅面结束都需要 关div -->
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− | <div class="clear extra_space"></div><!-- 一个章节结束需要clear来留白 如果觉得内容间差异大再加上extra_space -->
| + | <div class="clear"></div> |
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− | <h2>又来个章节 sis</h2>
| + | <h2> CaAP (Calcium Absorption Peptide)</h2> |
− | <div class="column full_size">
| + | <div class="column full_size"> |
− | <p>引入文字 ion.</p>
| + | <p>We hope that our project can solve the problem of calcium deficiency in the elderly. According to our investigation, due to the diet structure, the calcium absorption of Chinese people is not high, so we found CaAP from the perspective of improving the calcium absorption rate of the elderly.</p> |
− | <p>Firstly, the product is designed for oral use and our target user is the elderly. 英文分项不需要ol或ul</p>
| + | |
− | <ol>
| + | <p>We have successfully cloned our designed CaAP(Calcium Absorption Peptide) into pFAB plasmid and transformed the construct into <i>E. coli</i>. BL21 strain to demonstrate its expression.</p> |
− | <li>The produc</li>
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− | <li>The short pep</li>
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− | <li>The expression level of short peptides has a threshold, preventing any excessive calcium absorption that might lead to hypercalcemia 注意句子超长后的回折</li>
| + | <p>We chose to use a set of promoters to drive the expression of the specially designed CaAP(Calcium Absorption Peptide). Thus, the expression intensity of the gene of interest can be quantified and regulated.</p> |
− | <li>There is a kill swit.</li>
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− | </ol>
| + | <p>P1-P14 is a collection of precisely quantitated constitutive bacterial promoters that vary in strength and effectiveness. These promoters can be used to initiate the expression of the target gene in <i>E. coli</i> in a controlled, reliable way. </p> |
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| + | <p>To test the calcium-chelating ability of the designed CaAP, we purified it from the bacterial lysate of our successfully cloned strains(<i>E. coli</i>. BL21 transformed with pFAB-CaAP) using Ni-chelating affinity chromatography(CaAP contains an 6x-His tag at the C terminal). In this way, the effect of CaAP can be demonstrated through the differences of calcium ion concentration in the divalent calcium ion solution with or without CaAP.</p> |
| + | </div> |
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| + | <div class="clear"></div> |
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− | <p>
| + | <h2> Secretion peptides guided CaAP</h2> |
− | Based on relevant research, the product is exp 章节内分段 ina. tien China has an estimated 70 million osteoporosis patien China has an estimated 70 million osteoporosis patienChina has an estimated 70 million oste
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− | oporosis patienChina has an estimated 70 million osteoporosis pat
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− | </p>
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− | <p>Compared w 章节内分段 ique advantages tien China has an estimated 70 million osteoporosis patien China has an estimated 70 million osteoporosis patienChina has an estimated 70 million oste
| + | <div class="column full_size"> |
− | oporosis patienChina has an estimated 70 million osteoporosis pat tien China has an estimated 70 million osteoporosis patien China has an estimated 70 million osteoporosis patienChina has an estimated 70 million oste
| + | <p>To promote calcium bioavailability, CaAP needs to be expressed in our probiotics and subsequently secreted into the extracellular space. Therefore, we linked 5 signal peptides which belong to the Type II secretion system in <i>E. coli</i>., to the N terminal of CaAP, respectively. |
− | oporosis patienChina has an estimated 70 million osteoporosis pat tien China has an estimated 70 million osteoporosis patien China has an estimated 70 million osteoporosis patienChina has an estimated 70 million oste
| + | The 5 secretion peptides are listed below:</p> |
− | oporosis patienChina has an estimated 70 million osteoporosis pat.
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− | </p>
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| <table> | | <table> |
| <tr> | | <tr> |
− | <th> Header 1 这是igem hq的表格的写法 </th> <th> Header 2 </th> | + | <td> NSP4 </td> <td> <a href="http://parts.igem.org/Part:BBa_K3606042" target="_blank">BBa_K3606042</a> </td> |
| </tr> | | </tr> |
| <tr> | | <tr> |
− | <td> Content A 1 </td> <td> Content B 1 </td> | + | <td> OmpA </td> <td> <a href="http://parts.igem.org/Part:BBa_K3606043" target="_blank">BBa_K3606043</a> </td> |
| </tr> | | </tr> |
| <tr> | | <tr> |
− | <td> Content A 2 </td> <td> Content B 2 这是igem hq的表格的写法。表格都需要说明,之前之后都会有p来文字 </td> | + | <td> DsbA </td> <td> <a href="http://parts.igem.org/Part:BBa_K3606030" target="_blank">BBa_K3606030</a> </td> |
| </tr> | | </tr> |
− | </table>
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− |
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− | <p></p><!-- 无文字p是换行隔开,在关闭的html元素间回车是没有用的 -->
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− |
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− | <table class="striped"><!-- 这是另一个版本的表格,间隔行有阴影。一个页面不建议交叉两种格式。太宽表格可以加 responsive-table 方便窄屏 -->
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| <tr> | | <tr> |
− | <th>Name</th> <th>Item Name</th> <th>Item Price</th> | + | <td> PelB </td> <td> <a href="http://parts.igem.org/Part:BBa_K208004" target="_blank">BBa_K208004</a> </td> |
| </tr> | | </tr> |
| <tr> | | <tr> |
− | <td>Alvin</td> <td>Eclair</td> <td>$0.87</td> | + | <td> PhoA </td> <td> <a href="http://parts.igem.org/Part:BBa_K808028" target="_blank">BBa_K808028</a> </td> |
− | </tr>
| + | |
− | <tr>
| + | |
− | <td>Alan</td> <td>Jellybean</td> <td>$3.76</td>
| + | |
− | </tr> | + | |
− | <tr>
| + | |
− | <td>Jonathan</td> <td>Lollipop</td> <td>$7.00</td>
| + | |
| </tr> | | </tr> |
| </table> | | </table> |
| + | </div> |
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− | <p>Calciu 章节内分段 fety. tien China has an estimated 70 million osteoporosis patien China has an estimated 70 million osteoporosis patienChina has an estimated 70 million oste | + | <div class="column full_size"> |
− | oporosis patienChina has an estima
| + | <p>To test the secretion efficiency of our 5 secretion peptides fused CaAP(Calcium Absorption Peptide) guided by 5 secretion peptides, we cloned the Secretion peptide-CaAP segment into the pGEX vector respectively and transformed into <i>E. coli</i>. BL21 strain. Thus, quantified characterization of these secretion peptides fused with CaAP is expected to be achieved through the differences of calcium ion concentration in bacterial lysate and culture medium.</p></div> |
− | ted 70 million osteoporosis pat tien China has an estimated 70 million osteoporosis patien China has an estimated 70 million osteoporosis patienChina has an estimated 70 million oste
| + | |
− | oporosis patienChina has an estimated 70 mill
| + | |
− | ion osteoporosis pattien China has an estimated 70 million osteoporosis patien China has an estimated 70 million osteoporosis patienChina has an estimated 70 million oste | + | |
− | oporosis patienChina has an estimated 70 million osteoporosis pat
| + | |
− | </p>
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− | <p>From the us 章节内分段 etition.</p>
| + | <div class="row"> |
− | </div> | + | <div class="col m2 l3 hide-on-small-only"> </div> |
| + | <div class="col s12 m8 l6"> |
| + | <img src="https://static.igem.org/mediawiki/parts/6/67/T--Fudan--denoted_gel2_signalpeptides.png" alt="gel2 signal peptides" style="width:100%"> |
| + | <i>NSP4/PhoA/OmpA-CaAP expression with and without IPTG induction in the bacterial lysate.</i> |
| + | </div> |
| + | </div> |
| + | <div class="row"> |
| + | <div class="col m2 l3 hide-on-small-only"> </div> |
| + | <div class="col s12 m8 l6"> |
| + | <img src="https://static.igem.org/mediawiki/parts/0/0d/T--Fudan--File-T--Fudan--denoted_gel3_signalpeptides.png" alt="gel3 signal peptides" style="width:100%"> |
| + | <i>NSP4/PhoA/OmpA-CaAP expression expression with and without IPTG induction in the culture medium concentrated by TCA.</i> |
| + | </div> |
| + | </div> |
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− | <div class="clear"></div><!-- 一个章节结束需要clear来留白 -->
| + | <div class="clear"></div> |
| + | <h2> Mcb and quorum sensing </h2> |
| + | <div class="column full_size"> |
| + | <p>Due to the phenomenon that the elderly often forget to take medicine, we designed a quorum sensing module so that the flora can exist stably and maintain a certain amount in the intestinal. We also improved the relevant original parts as we found some problems with the system. </p> |
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− | <h2>再来例子 Market Analysis</h2>
| + | <p> McbABCDEFG is a full functional gene cluster that produces microcin B17 ( MccB17 ), a kind of antimicrobial peptide. In order to realize the regulation of its secretion, we divided it into two parts: McbABCD is an antibiotic coding gene cluster responsible for the maturation of MccB17. McbEFG is an immunity system coding gene cluster with efflux and immunity related protein. In our design, when the number of engineered bacteria is low, both McbABCD and McbEFG will be expressed, but when they are high, only McbABCD will be expressed. </p> |
− | <div class="column half_size">
| + | |
− | <p>文本在左侧一半,宽屏 According to the Chinese Dietetic Sociietary Nutrient Intakes of Chinese Residents", the calcium requirement for infants and young children increases with age, from 300mg/day to 800mg/day, and the calcium requirement for adolescents reaches 1000mg/day. The amof the required intake, with the small exception of some coastal and pastoral areas 图片在右侧一半</p>
| + | |
| | | |
− | <table> | + | <p> We have successfully cloned these two parts into pFAB plasmid. To be more detail, we used a series of constitutive promoters (P1/2/9/11/12/14) to drive McbABCD and PtetR to drive McbEFG, and place them in reverse on the same vector. We have also designed a plasmid with McbABCD (driven by P1/2/9/11/12/14) but without McbEFG in order to figure out the immune effect of McbEFG. These recombinant plasmids were transformed into BL21 for expression and their antibacterial effects were measured. </p> |
− | <tr>
| + | </div> |
− | <th> 这是igem hq的表格的写法 </th> <th> Header 2 </th> <th> Header 3 </th>
| + | <div class="row"> |
− | </tr> | + | <div class="col m2 l3 hide-on-small-only"> </div> |
− | <tr> | + | <div class="col s12 m8 l6"> |
− | <td> Content A 1 </td> <td> Content B 1 </td> <td> Content C 1 </td> | + | <img src="https://static.igem.org/mediawiki/2020/e/ea/T--Fudan--img_mcbabcdefg.png" alt="mcb ABCDEFG" style="width:100%"> |
− | </tr> | + | </div> |
− | <tr> | + | </div> |
− | <td colspan=2> 列融合 </td> <td> Content C 2 这是igem hq的表格的写法。半幅或者2/3幅可以用。<br/>行融合<br/>就<br/>用br了 </td> | + | <div class="column full_size"> |
− | </tr> | + | <p>In order to prove that McbABCDEFG does have an effective antibacterial effect as well as McbEFG can protect the engineered bacteria themselves and help the secretion of antimicrobial peptides more effectively, we designed the following experiment:</p> |
− | </table> | + | <p> We mixed WT <i>E. coli</i> (expressing GFP driven by plac) and <i>E. coli</i> with mcbABCD-mcbEFG-PtetR in different ratios (5:7 20:7 50:7), and measured the OD value of the bacteria two hours after adding an inducer, which can be used to reflect the antibacterial effect of antimicrobial peptides. We followed the same method as above to mix WT <i>E. coli</i> and <i>E. coli</i> with merely mcbABCD, induce and measure the OD value. </p> |
| | | |
− | </div> | + | <p>Our specific experimental steps are as follows: </p> |
− | <div class="column half_size"> | + | </div> |
− | <img src="https://static.igem.org/mediawiki/2020/0/00/T--Fudan--calcium_intake_graph.svg" alt="calcium intake graph"> | + | <div class="row"> |
− | </div>
| + | <div class="col m2 l2 hide-on-small-only"> </div> |
| + | <div class="col s12 m8 l8"> |
| + | <img src="https://static.igem.org/mediawiki/2020/d/d5/T--Fudan--img_flowchart1.png" alt="img_flowchart1" style="width:100%"> |
| + | </div> |
| + | </div> |
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− | <div class="clear extra_space"></div><!-- 一个章节结束需要clear来留白 因为马上最后一幅,建议extra_space -->
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− | <div class="column full_size">
| + | <div class="column full_size"> |
− | <p>每页结束需要小结,可以 ul ol 列表highlight,可以是每页第一段的另一种写法fety. tien China has an estimated 70 million osteoporosis patien China has an estimated 70 million osteoporosis patienChina has an estimated 70 million oste | + | <ol> |
− | oporosis patienChina has an estima
| + | <li> Culture WT(<i>E. coli</i> expressing GFP driven by plac), <i>E. coli</i> expressing mcbABCD (driven by p1\2\9\11\12\13) and <i>E. coli</i> expressing mcbABCD-mcbEFG-PtetR (driven by p1\2\9\11\12\13) separately at the same time for about</li> |
− | ted 70 million osteoporosis pat tien China has an estimated 70 million osteoporosis patien China has an estimated 70 million osteoporosis patienChina has an estimated 70 million oste
| + | <li> When the OD of WT reaches 0.6, mix WT with <i>E. coli</i> expressing driven by mcbABCD (driven by p1\2\9\11\12\13) or <i>E. coli</i> expressing mcbABCD-mcbEFG-PtetR (driven by p1\2\9\11\12\13), with ratio of 1:1, 4:1, 10:1. Induce them with IPTG(1mM/ml) for 2h. (The control group would be merely WT with IPTG and WT without IPTG). The inducer is used to make WT produce GFP. After mixture, immediately put the engineered <i>E. coli</i> on ice to cease the proliferation and measure their OD later for a precise mixture ratio</li> |
− | oporosis patienChina has an estimated 70 mi 每页结束需要小结,可以 ul ol 列表highlight,可以是每页第一段的另一种写法</p>
| + | <li> After induction, take 2mL of the culture to centrifugate the cell at 6000 rpm, 1 min</li> |
− | </div>
| + | <li> Wash with 500uL PBS then centrifugate again at 6000 rpm, 1 min </li> |
| + | <li> Fix with 400uL PFA for more than 10min, centrifugate again at 6000 rpm, 1 min </li> |
| + | <li> Re-suspend cells with 400uL PBS </li> |
| + | <li> Distribute into 96-well plates then use plate reader for further data collection</li> |
| + | </ol> |
| + | </div> |
| + | <div class="column half_size"> |
| + | <img src="https://static.igem.org/mediawiki/2020/1/16/T--Fudan--5mix7.jpeg" alt="IPTG 5mix7" /> |
| + | <h6>Figure 1. MEFL/particle for different Px and PxAD with the ratio of mixure (WT:Px=5:7) </h6></div> |
| + | |
| + | <div class="column half_size"> |
| + | <img src="https://static.igem.org/mediawiki/2020/b/b6/T--Fudan--20mix7.jpeg" alt="IPTG 20mix7" /> |
| + | <h6>Figure 2. MEFL/particle for different Px and PxAD with the ratio of mixure (WT:Px=20:7) </h6></div> |
| + | |
| + | <div class="column half_size"> |
| + | <img src="https://static.igem.org/mediawiki/2020/8/89/T--Fudan--50mix7.jpeg" alt="IPTG 50mix7" /> |
| + | <h6>Figure 3. MEFL/particle for different Px and PxAD with the ratio of mixure (WT:Px=50:7) </h6></div> |
| + | |
| + | |
| + | |
| + | <div class="clear"></div> |
| + | <div class="column full_size"> |
| + | <p> The result shows that <i>E. coli</i> with mcbABCD-mcbEFG-PtetR can more effectively inhibit the metabolism of other bacteria than those merely with mcbABCD in each group. With mcbEFG, <i>E. coli</i> are able to export antimicrobial peptides more effectively and supposed to have better immunity themselves so as to affect the surrounding WT’s metabolism(the expression of GFP), and better survive in the environment. While comparing to the control group, we can also see a significant decrease in the GFP expression result when there are <i>E. coli</i> with mcbABCD-mcbEFG-PtetR or mcbABCD, which indicates that the antimicrobial peptide (MccB17) encoded by mcbABCD does have a strong restraining effect on other microbial. That is to say, we can indeed increase the competitiveness of engineered bacteria by turning on the McbEFG gene, and turn off the McbEFG gene to block the antibacterial effect of antimicrobial peptides. </p> |
| + | |
| + | <p>It is worth noting that when the mixing ratio of WT <i>E. coli</i> and McbABCDEFG-expressing bacteria is 50:7, the OD values of each group are very close to those of the control group (only induced WT <i>E. coli</i>). This shows that when the concentration of the engineered bacteria is very low, the antimicrobial peptides are difficult to exert antibacterial effect and the engineered bacteria have no competitive advantage. This proves the necessity of our quorum sensing system. Because our engineered bacteria must first focus on their own growth and reach a certain number in order to effectively colonize the intestine. </p> |
| + | </div> |
| + | |
| + | <div class="row"> |
| + | <div class="col m2 l2 hide-on-small-only"> </div> |
| + | <div class="col s12 m8 l8"> |
| + | <img src="https://static.igem.org/mediawiki/2020/e/e9/T--Fudan--flowchart_kanamp.png" alt="flowchart kan amp" style="width:100%"> |
| + | </div> |
| + | </div> |
| + | |
| + | |
| + | <div class="column full_size"> |
| + | <p>We also designed experiments to measure the real-time antibacterial effects of McbABCDEFG and McbABCD. In this experiment, we gave WT and our engineered bacteria different antibiotic resistance, so through resistance screening, we can get both the number of WT and the number of engineered bacteria. We measure their OD value at different times after the two are mixed were mixed at a ratio of 1:1 or 1:4 to plotted their respective growth curves. It simulates the process of the engineering bacteria killing surrounding bacteria in the intestines. </p> |
| + | </div> |
| + | <div class="clear"></div> |
| + | |
| + | <h2>Measurement</h2> |
| + | |
| + | <div class="column full_size"> |
| + | <p>We hope for the old to make a full range of security for bone health. Based on the early part of the investigation, we found a lack of a proper way for the elderly to assess if their calcium status is healthy. Simultaneously, to evaluate whether our flora functions excessively and prevent absorbing too much calcium, we developed a measurement scheme - CaAsst. It provides for the elderly a parameter to assess their calcium levels - blood calcium concentration. We use the MTB as the key chemical to measurement the calcium. As the figure 1 blow shows, the experiment results show a linear relationship between calcium ion concentration and absorbance within a specific range. We also introduced a variety of interference simultaneously to show that the stability of this kind of measurement scheme. And there is also no apparent difference according to the figure 2. At the same time, CaAsst still has a lot of room for development, and in the future, it could even be widely promoted like a blood sugar test.</p> |
| + | </div> |
| + | <div class="column half_size"> |
| + | <img src="https://static.igem.org/mediawiki/2020/a/a3/T--Fudan--yicunanxian.png" alt="yicunanxian" /> |
| + | <h6>Figure 4. The fitting curve of 0-20mmol/L absorbance changes with calcium ions concentration, which conforms to Beale's law within this concentration range.</h6> |
| + | </div> |
| + | <div class="column half_size"> |
| + | <img src="https://static.igem.org/mediawiki/2020/8/87/T--Fudan--zongganrao.png" alt="zongganrao" /> |
| + | <h6>Figure 5. The curves of absorbance varying with the concentration of calcium ions and these curves are obtained from all experimental data</h6> |
| + | </div> |
| + | <div class="clear"></div> |
| + | |
| + | <h2> Entrepreneurship</h2> |
| + | <div class="column full_size"> |
| + | <p> Finally, we designed a thorough business plan for more financial support and promotion opportunities: from marketing research, product design to a feasible timeline for the future. We also consulted with experienced investors with great interest in our project. We communicated back and forth and signed a letter of intent to solidify future cooperation. And, you can get more information in <a href="/Team:Fudan/Entrepreneurship">https://2020.igem.org/Team:Fudan/Entrepreneurship</a>. |
| + | </p> |
| + | </div> |
| + | |
| + | <div class="clear"></div><!-- 一个章节结束需要clear来留白 因为马上最后一幅,建议extra_space --> |
| | | |
| <div class="column full_size"><div id="FudanSignature"> | | <div class="column full_size"><div id="FudanSignature"> |
− | <p>Signature: <a href="/Team:Fudan/Attributions#WeiZY" class="z-depth-2">WeiZY</a> | + | <p>Signature: <a href="/Team:Fudan/Attributions#WeiZY" class="z-depth-2">Ziyuan</a> |
| <a href="/Team:Fudan/Attributions#ZhangZR" class="z-depth-2">Zhenru</a> | | <a href="/Team:Fudan/Attributions#ZhangZR" class="z-depth-2">Zhenru</a> |
| + | <a href="/Team:Fudan/Attributions#ZhangJQ" class="z-depth-2">Jingqi</a> |
| + | <a href="/Team:Fudan/Attributions#ZhangGC" class="z-depth-2">Gaochen</a> |
| </p> | | </p> |
| </div></div> | | </div></div> |
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| <div class="container"><div class="row"> | | <div class="container"><div class="row"> |
| <div class="col m3 s12 row" | | <div class="col m3 s12 row" |
− | style="margin-top:4rem" > | + | style="margin-top:4rem" tabIndex="0" > |
| <a href="http://www.yfc.cn/" target="_blank"><img class="col s3 m6 l4 FudanSponsors" alt="Yunfeng Capital" src="https://static.igem.org/mediawiki/2020/b/ba/T--Fudan--YFcapital.svg"> </a> | | <a href="http://www.yfc.cn/" target="_blank"><img class="col s3 m6 l4 FudanSponsors" alt="Yunfeng Capital" src="https://static.igem.org/mediawiki/2020/b/ba/T--Fudan--YFcapital.svg"> </a> |
| <a href="http://www.fudan.edu.cn/en/" target="_blank"><img alt="Fudan University" class="col s3 m6 l4 FudanSponsors" src="https://static.igem.org/mediawiki/2020/9/9d/T--Fudan--sponsor2.svg"> </a> | | <a href="http://www.fudan.edu.cn/en/" target="_blank"><img alt="Fudan University" class="col s3 m6 l4 FudanSponsors" src="https://static.igem.org/mediawiki/2020/9/9d/T--Fudan--sponsor2.svg"> </a> |
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| <div class="col s12 l6 row"> | | <div class="col s12 l6 row"> |
| <div class="col s12 m4"> | | <div class="col s12 m4"> |
− | <span><a href="/Team:Fudan/Description#Project">Project</a></span> | + | <span tabIndex="2"><a href="/Team:Fudan/Description#Project">Project</a></span> |
| <ul><li><a href="/Team:Fudan/Description">Description</a></li> | | <ul><li><a href="/Team:Fudan/Description">Description</a></li> |
| <li><a href="/Team:Fudan/Sustainable">Sustainable</a></li> | | <li><a href="/Team:Fudan/Sustainable">Sustainable</a></li> |
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| </ul></div> | | </ul></div> |
| <div class="col s12 m4"> | | <div class="col s12 m4"> |
− | <span><a href="/Team:Fudan/Notebook">Notebook</a></span> | + | <span tabIndex="3"><a href="/Team:Fudan/Notebook">Notebook</a></span> |
| <ul><li><a href="/Team:Fudan/Safety">Safety</a></li> | | <ul><li><a href="/Team:Fudan/Safety">Safety</a></li> |
| <li><a href="/Team:Fudan/Poster">Poster</a></li> | | <li><a href="/Team:Fudan/Poster">Poster</a></li> |
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| </ul></div> | | </ul></div> |
| <div class="col s12 m4"> | | <div class="col s12 m4"> |
− | <span><a href="/Team:Fudan/Results">Results</a></span> | + | <span tabIndex="4"><a href="/Team:Fudan/Results">Results</a></span> |
| <ul><li><a href="/Team:Fudan/Contribution">Contribution</a></li><!-- replace Judging --> | | <ul><li><a href="/Team:Fudan/Contribution">Contribution</a></li><!-- replace Judging --> |
| <li><a href="/Team:Fudan/Implementation">Implementation</a></li> | | <li><a href="/Team:Fudan/Implementation">Implementation</a></li> |
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| <div class="col s12 l6 row"> | | <div class="col s12 l6 row"> |
| <div class="col s12 m4"> | | <div class="col s12 m4"> |
− | <span><a href="/Team:Fudan/Parts">Parts</a></span> | + | <span tabIndex="5"><a href="/Team:Fudan/Parts">Parts</a></span> |
| <ul><li><a href="/Team:Fudan/Parts#Basic">Basic part</a></li> | | <ul><li><a href="/Team:Fudan/Parts#Basic">Basic part</a></li> |
| <li><a href="/Team:Fudan/Parts#Composite">Composite part</a></li> | | <li><a href="/Team:Fudan/Parts#Composite">Composite part</a></li> |
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| </ul></div> | | </ul></div> |
| <div class="col s12 m4"> | | <div class="col s12 m4"> |
− | <span><a href="/Team:Fudan/Human_Practices">Outreach</a></span> | + | <span tabIndex="6"><a href="/Team:Fudan/Human_Practices">Outreach</a></span> |
| <ul><li><a href="/Team:Fudan/Human_Practices#Integrated">Integrated HP</a></li> | | <ul><li><a href="/Team:Fudan/Human_Practices#Integrated">Integrated HP</a></li> |
| <li><a href="/Team:Fudan/Education">Education</a></li> | | <li><a href="/Team:Fudan/Education">Education</a></li> |
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| </ul></div> | | </ul></div> |
| <div class="col s12 m4"> | | <div class="col s12 m4"> |
− | <span><a href="/Team:Fudan/Team">Team</a></span> | + | <span tabIndex="7"><a href="/Team:Fudan/Team">Team</a></span> |
| <ul><li><a href="/Team:Fudan/Team">Members</a></li> | | <ul><li><a href="/Team:Fudan/Team">Members</a></li> |
| <li><a href="/Team:Fudan/Attributions">Attributions</a></li> | | <li><a href="/Team:Fudan/Attributions">Attributions</a></li> |
| <li><a href="/Team:Fudan/Partnership">Partnership</a></li> | | <li><a href="/Team:Fudan/Partnership">Partnership</a></li> |
| <li><a href="/Team:Fudan/Acknowledgement">Acknowledgement</a></li> | | <li><a href="/Team:Fudan/Acknowledgement">Acknowledgement</a></li> |
− | <li><a href="https://2019.igem.org/Team:Fudan-TSI/Heritage" target="_blank">Heritage</a></li> | + | <li><a href="/Team:Fudan/Heritage">Heritage</a></li> |
| </ul></div> | | </ul></div> |
| <div class="col s12 m4"> </div> | | <div class="col s12 m4"> </div> |