Difference between revisions of "Team:OUC-China/Human Practices"

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<h3 class="text-center"><img src="https://static.igem.org/mediawiki/2016/c/cf/T--OUC-China--head-icon1.fw.png">Background<img src="https://static.igem.org/mediawiki/2016/f/f8/T--OUC-China--head-icon2.fw.png"></h3>
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<h3 class="text-center"><img src="https://static.igem.org/mediawiki/2016/c/cf/T--OUC-China--head-icon1.fw.png" alt="icon">Overview<img src="https://static.igem.org/mediawiki/2016/f/f8/T--OUC-China--head-icon2.fw.png" alt="icon"></h3>
<p>Nowadays,quantitative has become a general trend, and of which the control of gene expression play a vital role. Until now, there’re several ways to control gene expression and consequently achieve stoichiometry and functional protein products.</p>
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<p>This year our team devoted to a novel regulation element, which makes great sense in synthetic biology but may be a little obscure for those who are unfamiliar with this field. Our regulation element works out through synthetic biology and then being acknowledged by the public. So our work was performed around three cores: our design, synthetic biology and public acknowledgement. And below are the diagrams of our project working mechanism.</p>
<p>Regulation of gene expression includes a wide range of mechanisms that are used by cells to increase or decrease the production of specific gene  products(protein or RNA). Virtually any step of gene expression can be modulated, from transcriptional initiation, to RNA processing, and to the post-translational modification of a protein.</p>
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<img src="https://static.igem.org/mediawiki/2016/2/2b/T--OUC-China--hp-0.jpg" alt="3 parts of our human practices" />
                                <b style="font-size:18px">Modification of DNA(左侧插图~~~~)</b>
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<p>We carried out our human practices in three parts:</p>
                                <p>In eukaryotes, the modification of DNA’s chromatin structure, such as histone modifications directed by DNA methylation[1], ncRNA, or DNA-binding protein, may up or down regulate the expression of a gene.</p>
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<p><b>1. Communicating and Improving: </b>Consulted experts of synthetic biology to evaluate our project and improved our design. <br><b>2. Investigating and Promoting:</b> Investigated the popularity and acknowledgement of our design as well as synthetic biology and devoted to promoting them.<br><b>3. Potential Application:</b> Did some practical research and looked for potential specific proteins that can be applied with our design.</p>
                                <b style="font-size:18px">Regulation of transcription(左侧插图~~~~)</b>
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                                <p>This controls when transcription occurs and the amount of RNA be created. The mechanisms usually includes specificity factors, general transcription factors, repressors, activators, enhancers [2] and silencers.</p>
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                                <b style="font-size:18px">Post-transcriptional regulation [3] (左侧插图~~~~)</b>
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                                <p>In eukaryotes, there have some mechanisms on how much the mRNA is translated into proteins [4]. Cells do this by modulating the capping, splicing, addition of a Poly(A) Tail, the sequence-specific nuclear export rates, and, in several contexts, sequestration of the RNA transcript.</p>
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                                <b style="font-size:18px">Regulation of translation(左侧插图~~~~)</b>
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                                <p>The translation of mRNA can also be controlled by a number of mechanisms[5], mostly at the level of initiation. The secondary structure of mRNA, antisense RNA binding, or protein binding[6] can all modulate the recruitment of the small ribosomal subunit. </p>
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                                <p>So many regulation methods have developed to achieve gene expression on desired level. Accurate as they are, they can hardly control the relative expression of several cistrons simultaneously. They are usually performed on operon level and may not have difference influence on each cistron.</p>
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                                <p>But on certain condition, it’s of vital importance to realize differential gene expression in polycistrons. For example, <a href="https://2014.igem.org/Team:Imperial">Team Imperial 2014</a> aimed to biosynthesize bacterial cellulose in <i>E.coil</i>, they need to transform two cistrons of different expression level into <i>E.coil</i>. Without efficient approaches, they did lots of jobs. They selected proper copied plasmid among 9 plasmids and then measured 15 Anderson promotors of different strength and finally selected a proper combination for the two different cistrons. </p>
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                                <p>This year, OUC-iGEM team devoted to exploring a novel regulation method on post-transcriptional level to realize differential expression in a polycistron. Details see the design part.</p>
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<h3 class="text-center"><img src="https://static.igem.org/mediawiki/2016/c/cf/T--OUC-China--head-icon1.fw.png">Design<img src="https://static.igem.org/mediawiki/2016/f/f8/T--OUC-China--head-icon2.fw.png"></h3>
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<h3 class="text-center">PartⅠ</h3>
                                <b style="font-size:18px">Overview</b>
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<h3 class="text-center"><img src="https://static.igem.org/mediawiki/2016/c/cf/T--OUC-China--head-icon1.fw.png" alt="icon">Communicating and Improving<img src="https://static.igem.org/mediawiki/2016/f/f8/T--OUC-China--head-icon2.fw.png" alt="icon"></h3>
<p>In C.Cellulolyticum,it is reported that a novel regulation method was employed to regulate the differential expression of multiple cistrons within the same operon[1]. And we came out the idea of exploiting this mechanism to a toolkit using for regulating differential and quantitative expression within a polycistron.</p>
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<p>At the beginning of our project design, we met a tough problem of choosing chassis. We had two choices, <i>B.subtilis</i> and <i>E.coli</i>. The <i>B.subtilis</i> is a kind of Gram-positive bacterium and is more similar to our prototype---<i>C.cellulolyticum</i>, but the <i>E.coli</i> is more versatile and widely used. </p>
                                <p>The principle is as follows:</p>
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<p>We summarized our doubts and consulted Dr. Xu Chenggang, an expert in <i>C.cellulolyticum</i> research. Then we got the advice of comparing more differences that related to our design and he recommended us several useful software. In the process of blasting and comparing enzyme system between the two, we found that <i>E.coli</i> has no exoribonuclease that can digest from 5’ termini, which will be of great benefit to our whole design. With the help of expert and our deep research, we made sure our chassis and went on with further design.</p>
<img src="https://static.igem.org/mediawiki/2016/6/62/T--OUC-China--project-2.1.png" class="img-responsive">
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<img src="https://static.igem.org/mediawiki/2016/9/9a/T--OUC-China--hp1.jpg" class="img-responsive" alt="2016 OUC-iGEMers with Dr. Xu">
                                <p class="text-center" style="font-size:14px;">Figure 1. Diagram of the post-transcriptional regulation method</p>
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                <p class="text-center" style="font-size:14px;">Figure 1. 2016 OUC-iGEMers with Dr. Xu</p>
                                <p>In bacteria genomes, genes are frequently organized as an operon that mostly encodes protein complexes, which is an efficient means to regulate transcription of multiple genes simultaneously. It is firstly transcribed by its sole promoter into the primary mRNA.<br>And then, in <i>E.coli</i> cases, the processing is initiated via a primary cleavage by RNase E[2]. It is a single-stranded, nonspecific endonuclease with preference for cleaving A/U-rich sequence and its cleavage signals locate in the intergenic regions. Primary transcript then will be split by RNase E into several secondary transcripts. [Fig.3]<br>Stability of these secondary transcripts varied widely due to their distinct terminal sequences that convey resistance to exonuclease degradation. In other words, stem loops with lower free energy can protect mRNA more from exoribonuclease degradation and stay longer in cells, thus the transcripts can  translate more proteins. <br>With this system, we can regulate multiple genes in one operon express on desired quantitative level, realize multiple cistrons express coordinately, just like a melodic cistrons concerto.<br>To achieve this, we constructed dual-fluorescent and tri-fluorescent reporter system and designed a series of stem-loops to insert into the reporter system to test this regulating method. Following are our basal designs. </p>
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<p>Then in September, we joined the CCiC (Conference of China iGEMer Committee) held by SYSU, which is a communicating platform for Chinese iGEM teams. About 30 teams and 300 iGEMers from all over China gathered together. We shared our design with other iGEMers and we were glad that we received many specific responses, from constructive project design to detailed data process skills. Then according to those suggestions, we modified our original design and added two supplementary proof experiments, which was essential to the integrity of our project.</p>
                                <b style="font-size:18px">Dual-fluorescent reporter system</b>
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<img src="https://static.igem.org/mediawiki/2016/f/f4/T--OUC-China--hp2.jpg" class="img-responsive" alt="Open ceremony of CCIC" />
<p>We want to test the protection effect on the upstream gene from stem-loops, thus we primarily construct a dual-fluorescent reporter system. The circuit is as follows: </p>
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                <p class="text-center" style="font-size:14px;">Figure 2.Open ceremony of CCIC</p>
<img src="https://static.igem.org/mediawiki/2016/e/e1/T--OUC-China--project-2.2.png" class="img-responsive">
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                                <p class="text-center" style="font-size:14px;">Figure 2. Dual-fluorescent reporter</p>
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                                <p>The promoter is inducible with arabinose and the two fluorescent reporter gfp and mCherry with their RBS are two parts in the iGEM part library, numbered <a href="http://parts.igem.org/Part:BBa_I13500">BBa_I13500</a> and <a href="http://parts.igem.org/Part:BBa_J06602">BBa_J06602</a>, the RNase E site between them is from <i>E.coli</i> pap operon [3], a relative high efficient endonuclease cleavage site through experiments. The stem-loops are from the series of stem loops we designed.</p>
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                                <b style="font-size:18px">Stem-loops</b>
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<p>We designed a series of stem-loops of gradient free energy to test this regulation method. And we determined the free energy of stem-loops with RNA folding program (Mfold). [4][5]</p>
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                                <p>We designed stem-loops in the view of 3 parts: loop, stem and a tail ahead with a pair of CG as "sealing nucleotides", see as figure 3.</p>
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<img src="https://static.igem.org/mediawiki/2016/8/84/T--OUC-China--project-2.3.png" class="img-responsive">
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                                <p class="text-center" style="font-size:14px;">Figure 3: Diagram of the designed stem-loops</p>
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                                <p>Loop: Since the loop makes little contribution to the whole free energy, we used the same loop throughout our designs. That is U-U-A-C-A-U-G-A-U-U. <br>Stem: The stem-loop's free energy is mainly decided by the pairing stem section. We chose the ratio of "A+U" to "C+G" as 1:1. For too many C/G may cause the energy too low and hard to adjust, whereas too many A/U are easy to cause terminating effect owing to the structure similarity to rho-independent terminators[6].. Choosing the ratio as 1:1, we can easily adjust the free energy by change the length of the paring stem. <br>"Sealing nucleotides": A pair of CG at the end of a stem-loop can help stabilizing the secondary structure by avoiding flanking extra base paring in different context circuit. <br>Following these principles, the stem-loops we designed are diaplayed in table 1, they are of gradient free energy:
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                                <p>We measured these stem loops in the dual-fluorescent reporter system. By analyzing the amount of mRNA and protein expressed in different circuits, we aimed to explore the correlation between the folding free energy and the quantitative expression. We measured the quantitative expression on two levels: the transcriptional level and the translational level. On the transcriptional level, we aimed to find if the difference is actually caused by the varied mRNA degradation. Upon the translational level, we explored the possibility and prospect of this post-transcriptional regulation mechanism on practical application.</p>
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                                <b style="font-size:18px">Tri-fluorescent reporter system</b>
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                                <p>By the dual-fluorescent reporter system we obtain the correlation between  folding free energy and quantitative expression of mRNA and protein fluorescence. Then we constructed a tri-fluorescent reporter system to furtherly test this relationship. The circuit is as follows:</p>
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                                <p class="text-center" style="font-size:14px;">Figure 4</p>
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                                <p>We use tri-fluorescent reporters to test the correlation we obtained, the inserted CFP with RBS is also a part in the part library, numbered <a href="http://parts.igem.org/Part:BBa_I13600">BBa_I13600.</a> <br>We use this circuit to measure the effect of two different stem loops, with  folding free energy of -51.4kcal/mol and -25.6kcal/mol. Detailed data see our <a href="https://2016.igem.org/Team:OUC-China/Results">result page</a>.</p>
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                                <b style="font-size:18px">Future</b>
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                                <p>This year, our team devoted to exploring a novel regulation method on the post-transcription level. Once the correlation is established and confirmed, we can apply this to tune the expression of polycistron of interest. To achieve our goals, we have 3 steps to go: </p>
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                                <p>a)Through our data analysis and modelling get the correlation. This step is finished in our lab with the basal circuit, the constructed fluorescent reporter system. <br>b)To apply this regulation method to practice using, we will develop it as a toolkit. Now we are exploiting a software to realize structure prediction base on the different folding free energy according to the target ratio.<br>c)Then we can employ our software a specific protein for expression optimizing. This can be used for industrial fermentation, so we are consulting specialists and connecting with related industries for advice.</p>
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<h3 class="text-center"><img src="https://static.igem.org/mediawiki/2016/c/cf/T--OUC-China--head-icon1.fw.png">Reference<img src="https://static.igem.org/mediawiki/2016/f/f8/T--OUC-China--head-icon2.fw.png"></h3>
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                                <b style="font-size:18px">Ref. in Background:</b>
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<h3 class="text-center">Part Ⅱ</h3>
                                <p style="font-size:14px">[1].Bell JT, Pai AA, Pickrell JK, Gaffney DJ, Pique-Regi R, Degner JF, Gilad Y, Pritchard JK (2011). "DNA methylation patterns associate with genetic and gene expression variation in HapMap cell lines". Genome Biology. 12 (1): <br>[2]. Austin S, Dixon R (Jun 1992). "The prokaryotic enhancer binding protein NTRC has an ATPase activity which is phosphorylation and DNA dependent". The EMBO Journal. 11(6): 2219–28. <br>[3].Weaver, Robert J. (2007). "Part V: Post-transcriptional events". Molecular Biology. Boston: McGraw Hill Higher Education..<br>[4].Hu W, Coller J (2012). "What comes first: translational repression or mRNA degradation? The deepening mystery of microRNA function". Cell Res. 22 (9): 1322–4. doi:10.1038/cr.2012.80<br>[5].Kozak M (1999). "Initiation of translation in prokaryotes and eukaryotes". Gene. 234 (2): 187–208. doi:10.1016/S0378-1119(99)00210-3. PMID 10395892<br>[6].Malys N, McCarthy JEG (2010). "Translation initiation: variations in the mechanism can be anticipated". Cellular and Molecular Life Sciences. 68 (6): 991-1003. doi: 10.1007/s00018-010-0588-z.</p>
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<h3 class="text-center"><img src="https://static.igem.org/mediawiki/2016/c/cf/T--OUC-China--head-icon1.fw.png" alt="icon">Investigating and Promoting<img src="https://static.igem.org/mediawiki/2016/f/f8/T--OUC-China--head-icon2.fw.png" alt="icon"></h3>
                                <b style="font-size:18px">Ref. in Design:</b>
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<p>Our novel <b>regulation element</b> reflects on synthetic biology as the realization of quantitative expression. To explore the potential promotion of our design, we firstly carried out a questionnaire survey about <b>quantification and synthetic biology</b>.</p>
                                <p style="font-size:14px">[1] Xu, C., Huang, R., Teng, L., Jing, X., Hu, J., Cui, G., . . . Xu, J. (2015). Cellulosome stoichiometry in Clostridium cellulolyticum is regulated by selective RNA processing and stabilization. Nat Commun, 6, 6900. doi: 10.1038/ncomms7900.<br>[2] Carpousis, A. J., Luisi, B. F., & McDowall, K. J. (2009). Endonucleolytic initiation of mRNA decay in Escherichia coli. Progress in molecular biology and translational science, 85, 91-135.<br>[3]Bricker, A. L., & Belasco, J. G. (1999). Importance of a 5′ Stem-Loop for Longevity ofpapA mRNA in Escherichia coli. Journal of bacteriology, 181(11), 3587-3590.<br>[4] Zuker M. Mfold web server for nucleic acid folding and hybridization prediction[J]. Nucleic acids research, 2003, 31(13): 3406-3415.<br><a href="http://unafold.rna.albany.edu/?q=mfold/RNA-Folding-Form">[5] http://unafold.rna.albany.edu/?q=mfold/RNA-Folding-Form</a><br>[6] Yarnell W S, Roberts J W. Mechanism of intrinsic transcription termination and antitermination[J]. Science, 1999, 284(5414): 611-615.</p>
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<p>We analyzed the survey result, to our astonishment, most people were lack of basic perception about quantification. For example, among the more than 500 investigators, 40% people didn’t consider meals combination as an example of quantification, and more than half frankly said that they knew little about synthetic biology. <a href="https://static.igem.org/mediawiki/2016/0/02/T--OUC-China--questionaire-result.pdf">Click here</a> to see the survey result.</p>
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<img src="https://static.igem.org/mediawiki/2016/d/d7/T--OUC-China--hp3.jpg" class="img-responsive" alt="Public Knowledge of Synthetic Biology">
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                <p class="text-center" style="font-size:14px;">Figure 3  extent of the public knowing of synthetic biology</p>
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<p>Given the insufficient popularization of quantification as well as synthetic biology, we focused on the spreading of basal knowledge. From campus to society, we used as much resource as possible to spreading synthetic biology and quantification.</p>
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<p style="font-size:20px;"><b>StepⅠ Spreading around us</b></p>
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<p>For people who are interested in nature science like university students, we designed series of activities for them to enjoy personal experience about synthetic biology.<br> Firstly, in our school, we held lectures and attracted about 100 students to introduce synthetic biology in details and got the feedback that they were interested in the newly developed subject and some were grouped together to explore more about it. (Figure 4)<br>Then, we held an academic summer camp and invited about 20 students from different universities. We stayed together with them for two weeks and shared a lot with them. Besides basic communicating in literature and experiment skills, we also held a small academic jamboree activity, with integral presentation and poster sections. (figure 5) Moreover, after the camp closing, we received the feedback that they enjoyed this academic exploration very much and would share what they had learned with more people.</p>
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<p class="text-center" style="font-size:14px;">Figure4. We held lectures in our school</p>
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<img src="https://static.igem.org/mediawiki/2016/6/61/T--OUC-China--hp5.jpg" class="img-responsive" alt="Students were presenting their posters">
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<p class="text-center" style="font-size:14px;">Figure5. Students were presenting their posters</p>
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<p style="font-size:20px;"><b>StepⅡ Popularizing to the public</b></p>
 +
<p>For the public, we tried to publicize synthetic biology by using relatively easy words toward as many people as possible through our activities.<br> We went to communities and tourist attractions and met people from the young to the old. With the help of our distributed brochures, we explained basic knowledge of synthetic biology and the meaning of quantification in their daily lives. (Figure 6)<br>We also posted our original video on website which used simple words and vivid flash to introduce the basic knowledge of synthetic biology. (Figure 7)</p>
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<img src="https://static.igem.org/mediawiki/2016/2/29/T--OUC-China--hp6.jpg" class="img-responsive" alt="Popularizing to the public">
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<p class="text-center" style="font-size:14px;">Figure6. Popularizing to the public</p>
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<div class="col-md-6">
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<img src="https://static.igem.org/mediawiki/2016/b/bb/T--OUC-China--hp-7.jpg" class="img-responsive" alt="Popularizing to the public">
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<p class="text-center" style="font-size:14px;">Figure7. Popularizing to the public</p>
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</div>
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</div>
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<p style="font-size:20px;"><b>StepⅢ Further communicating via social service organization</b></p>
 +
<p>Other novel ways are come up to spread synthetic besides above traditional ways. We got connected with Qingdao Association for Science and Technology, a non-profit social service organization for popularizing science and technology knowledge to the public<br> At there, we got the chance to displayed scientific knowledge to visitors. For example, we held an academic lecture for a delegation from Tibet, Autonomous Region, where the economic and educational development is relatively backward (Figure 8). According to the feedback from them, they were willing to know more innovative things and encourages us to do more.</p>
 +
<img src="https://static.igem.org/mediawiki/2016/4/45/T--OUC-China--hp-8.jpg" class="img-responsive" alt="We held a lecture to the delegation from Tibet">
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<p class="text-center" style="font-size:14px;">Figure8. We held a lecture to the delegation from Tibet</p>
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<h3 class="text-center"><img src="https://static.igem.org/mediawiki/2016/c/cf/T--OUC-China--head-icon1.fw.png" alt="icon">Youth education<img src="https://static.igem.org/mediawiki/2016/f/f8/T--OUC-China--head-icon2.fw.png" alt="icon"></h3>
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<p>For people who are more interested in nature science like university students, we designed series activities for them to enjoy personal experience about syntheticbiology. We held a summer camp and invited about 30 students from different universities all over China, like Shandong Agricultural University etc. By holding jamboree to present posters, doing experiment of molecular biology, doing literature based on research, students enjoyed themselves in synthetic biology and achieved necessary skill for academic exploration. In our school, we also held similar academic activities to attract more potential students to join in synthetic biology. For example, we held lectures for about 100 students to introduce synthetic biology in details.</p>
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<div class="row">
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<img src="https://static.igem.org/mediawiki/2016/d/d7/T--OUC-China--hp3.jpg" class="img-responsive">
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<div class="col-md-6">
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<img src="https://static.igem.org/mediawiki/2016/d/dd/T--OUC-China--hp4.jpg" class="img-responsive">
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<div class="clearfix"></div>
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<hr>
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<br id="float05">
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<h3 class="text-center"><img src="https://static.igem.org/mediawiki/2016/c/cf/T--OUC-China--head-icon1.fw.png" alt="icon">Consulting with specialists<img src="https://static.igem.org/mediawiki/2016/f/f8/T--OUC-China--head-icon2.fw.png" alt="icon"></h3>
 +
<p>When designing our project in April, we went to visited researcher Chengang, Xu in Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences. At that time, we wanted to have the feasibility analysis of our design. More importantly, we met problems about chassis. We initially design our project based on B.subtilis, a kind of gram-positive bacterium but after consulting Professor Xu, who did research related to our project and suggested us to use <i>E.coli</i>, which would make our experiment more feasible. We then compared the enzyme system both in B.subtilis and <i>E.coli</i> and found that in <i>E.coli</i> there was no enzyme cleavage from 5’. Therefore, we finally determined to use <i>E.coli</i>, the more versatile chassis.</p>
 +
<img src="https://static.igem.org/mediawiki/2016/6/61/T--OUC-China--hp5.jpg" class="img-responsive">
 +
<p>In September, we joined the CCiC( Conference of China iGEMer Committee), which was a platform for Chinese iGEM teams to communicate with each other. About 30 teams and 300 iGEMers from all over China gathered together this year and we got numerous responses from others, including suggestions on the thoughts of experiments, heated discussions on pathway design, and reflections on safety and application. Attendance of CCiC helped us acquire both reflections and inspiration from communicating, and improved our projects in the end.</p>
 +
<img src="https://static.igem.org/mediawiki/2016/2/29/T--OUC-China--hp6.jpg" class="img-responsive">
 +
<p>Particularly, we consulted our project and experiment with Haoqian, Zhang, the co-founder of an iGEM startup in China named Bluepha. He was the former leader of Peking iGEM 2010 and had rich experiment of iGEM. At the beginning, he thought quantification had the significant meaning in synthetic biology as well. He then gave us lots of specific and useful suggestion. For example, he advised us to exchange the upstream and downstream proteins to make our project more powerful. We adopted this suggestion and modified our project when we came back from CCiC. Besides, we met problems with our modeling. Another former leader of OUC iGEM, Yang, Liu suggested us to solve these problems in other alternative ways, which would make our work more efficient. </p>
 
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<p style="margin-top:20px;">Thanks to:<img src="https://static.igem.org/mediawiki/2016/5/57/T--OUC-China--foot1.jpg" style="margin-left:10px;" alt="Qingdao Institute of Bioenergy and Bioprocess Technology,Chinese Academy of Sciences"><img src="https://static.igem.org/mediawiki/2016/f/f0/T--OUC-China--foot2.jpg" style="margin-left:10px;" alt="Biolabs"></p>
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<p style="margin-top:20px;">Designed and built by @ Jasmine Chen and @ Zexin Jiao</p>
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<p style="margin-top:20px;">Code licensed under Apache License v4.0</p>
 
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<p>Thanks to:<i><img src="image/ouc-footer-1.fw.png"></i><i><img src="image/ouc-footer-2.fw.png"></i></p>
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<p style="margin-top:20px;">E-mail: oucigem@163.com</p>
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<p>Follow us on Facebook@ iGEM OUC<img src="https://static.igem.org/mediawiki/2016/9/94/T--OUC-China--foot3.png" style="margin-left:20px;" alt="Facebook"></p>
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<p>We are OUC-iGEM<img src="https://static.igem.org/mediawiki/2016/5/58/T--OUC-China--foot4.png" style="margin-left:10px;" alt="logo-one"><img src="https://static.igem.org/mediawiki/2016/9/9b/T--OUC-China--foot5.png" style="margin-left:10px;" alt="logo-two"></p>
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Revision as of 09:18, 14 October 2016

Human practices


iconOverviewicon

This year our team devoted to a novel regulation element, which makes great sense in synthetic biology but may be a little obscure for those who are unfamiliar with this field. Our regulation element works out through synthetic biology and then being acknowledged by the public. So our work was performed around three cores: our design, synthetic biology and public acknowledgement. And below are the diagrams of our project working mechanism.

3 parts of our human practices

We carried out our human practices in three parts:

1. Communicating and Improving: Consulted experts of synthetic biology to evaluate our project and improved our design.
2. Investigating and Promoting: Investigated the popularity and acknowledgement of our design as well as synthetic biology and devoted to promoting them.
3. Potential Application: Did some practical research and looked for potential specific proteins that can be applied with our design.



PartⅠ

iconCommunicating and Improvingicon

At the beginning of our project design, we met a tough problem of choosing chassis. We had two choices, B.subtilis and E.coli. The B.subtilis is a kind of Gram-positive bacterium and is more similar to our prototype---C.cellulolyticum, but the E.coli is more versatile and widely used.

We summarized our doubts and consulted Dr. Xu Chenggang, an expert in C.cellulolyticum research. Then we got the advice of comparing more differences that related to our design and he recommended us several useful software. In the process of blasting and comparing enzyme system between the two, we found that E.coli has no exoribonuclease that can digest from 5’ termini, which will be of great benefit to our whole design. With the help of expert and our deep research, we made sure our chassis and went on with further design.

2016 OUC-iGEMers with Dr. Xu

Figure 1. 2016 OUC-iGEMers with Dr. Xu

Then in September, we joined the CCiC (Conference of China iGEMer Committee) held by SYSU, which is a communicating platform for Chinese iGEM teams. About 30 teams and 300 iGEMers from all over China gathered together. We shared our design with other iGEMers and we were glad that we received many specific responses, from constructive project design to detailed data process skills. Then according to those suggestions, we modified our original design and added two supplementary proof experiments, which was essential to the integrity of our project.

Open ceremony of CCIC

Figure 2.Open ceremony of CCIC



Part Ⅱ

iconInvestigating and Promotingicon

Our novel regulation element reflects on synthetic biology as the realization of quantitative expression. To explore the potential promotion of our design, we firstly carried out a questionnaire survey about quantification and synthetic biology.

We analyzed the survey result, to our astonishment, most people were lack of basic perception about quantification. For example, among the more than 500 investigators, 40% people didn’t consider meals combination as an example of quantification, and more than half frankly said that they knew little about synthetic biology. Click here to see the survey result.

Public Knowledge of Synthetic Biology

Figure 3 extent of the public knowing of synthetic biology

Given the insufficient popularization of quantification as well as synthetic biology, we focused on the spreading of basal knowledge. From campus to society, we used as much resource as possible to spreading synthetic biology and quantification.

StepⅠ Spreading around us

For people who are interested in nature science like university students, we designed series of activities for them to enjoy personal experience about synthetic biology.
Firstly, in our school, we held lectures and attracted about 100 students to introduce synthetic biology in details and got the feedback that they were interested in the newly developed subject and some were grouped together to explore more about it. (Figure 4)
Then, we held an academic summer camp and invited about 20 students from different universities. We stayed together with them for two weeks and shared a lot with them. Besides basic communicating in literature and experiment skills, we also held a small academic jamboree activity, with integral presentation and poster sections. (figure 5) Moreover, after the camp closing, we received the feedback that they enjoyed this academic exploration very much and would share what they had learned with more people.

We held lectures in our school

Figure4. We held lectures in our school

Students were presenting their posters

Figure5. Students were presenting their posters

StepⅡ Popularizing to the public

For the public, we tried to publicize synthetic biology by using relatively easy words toward as many people as possible through our activities.
We went to communities and tourist attractions and met people from the young to the old. With the help of our distributed brochures, we explained basic knowledge of synthetic biology and the meaning of quantification in their daily lives. (Figure 6)
We also posted our original video on website which used simple words and vivid flash to introduce the basic knowledge of synthetic biology. (Figure 7)

Popularizing to the public

Figure6. Popularizing to the public

Popularizing to the public

Figure7. Popularizing to the public

StepⅢ Further communicating via social service organization

Other novel ways are come up to spread synthetic besides above traditional ways. We got connected with Qingdao Association for Science and Technology, a non-profit social service organization for popularizing science and technology knowledge to the public
At there, we got the chance to displayed scientific knowledge to visitors. For example, we held an academic lecture for a delegation from Tibet, Autonomous Region, where the economic and educational development is relatively backward (Figure 8). According to the feedback from them, they were willing to know more innovative things and encourages us to do more.

We held a lecture to the delegation from Tibet

Figure8. We held a lecture to the delegation from Tibet



iconYouth educationicon

For people who are more interested in nature science like university students, we designed series activities for them to enjoy personal experience about syntheticbiology. We held a summer camp and invited about 30 students from different universities all over China, like Shandong Agricultural University etc. By holding jamboree to present posters, doing experiment of molecular biology, doing literature based on research, students enjoyed themselves in synthetic biology and achieved necessary skill for academic exploration. In our school, we also held similar academic activities to attract more potential students to join in synthetic biology. For example, we held lectures for about 100 students to introduce synthetic biology in details.



iconConsulting with specialistsicon

When designing our project in April, we went to visited researcher Chengang, Xu in Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences. At that time, we wanted to have the feasibility analysis of our design. More importantly, we met problems about chassis. We initially design our project based on B.subtilis, a kind of gram-positive bacterium but after consulting Professor Xu, who did research related to our project and suggested us to use E.coli, which would make our experiment more feasible. We then compared the enzyme system both in B.subtilis and E.coli and found that in E.coli there was no enzyme cleavage from 5’. Therefore, we finally determined to use E.coli, the more versatile chassis.

In September, we joined the CCiC( Conference of China iGEMer Committee), which was a platform for Chinese iGEM teams to communicate with each other. About 30 teams and 300 iGEMers from all over China gathered together this year and we got numerous responses from others, including suggestions on the thoughts of experiments, heated discussions on pathway design, and reflections on safety and application. Attendance of CCiC helped us acquire both reflections and inspiration from communicating, and improved our projects in the end.

Particularly, we consulted our project and experiment with Haoqian, Zhang, the co-founder of an iGEM startup in China named Bluepha. He was the former leader of Peking iGEM 2010 and had rich experiment of iGEM. At the beginning, he thought quantification had the significant meaning in synthetic biology as well. He then gave us lots of specific and useful suggestion. For example, he advised us to exchange the upstream and downstream proteins to make our project more powerful. We adopted this suggestion and modified our project when we came back from CCiC. Besides, we met problems with our modeling. Another former leader of OUC iGEM, Yang, Liu suggested us to solve these problems in other alternative ways, which would make our work more efficient.




Cistrons Concerto

About:

Thanks to:Qingdao Institute of Bioenergy and Bioprocess Technology,Chinese Academy of SciencesBiolabs

Designed and built by @ Jasmine Chen and @ Zexin Jiao

Code licensed under Apache License v4.0

Contact us:

E-mail: oucigem@163.com

Follow us on Facebook@ iGEM OUCFacebook

Find us on Google Map

We are OUC-iGEMlogo-onelogo-two