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− | <h4>At dawn of time, the universe was like a gigantic egg and there was nothing but chaos.</h4>
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− | <h1 class="post-title">E. pangu: The Pioneer of Mars</h1> | + | <h1 class="post-title">Crispr And Verification</h1> |
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− | <h2 class="back"><strong>Background</strong></h2> | + | <h2 class="back"><strong>Crispr</strong></h2> |
− | <p>Since the onset of industrial era, the environment on earth keeps getting worse day by day because of the burning of fossil fuel and continuous deforestation. Recently, the discovery of Kepler-452b publicly announced by NASA calls public attention to space immigration again. As a matter of fact, the immigration to Mars, the most earthlike planet in solar system, still should be considered as priority. </p> | + | <h3 class="back"><strong>Introduction</strong></h3> |
− | <p>Before Mars immigration for human beings, it is necessary to send pioneer organism there to assess the feasibility of human immigration. What’s more, special pioneer organism can be also used to change the Martian atmosphere to make it more suitable for other organisms. The first three gaseous components of Martian atmosphere are carbon dioxide (96%), argon (1.93%) and nitrogen (1.89%)[1]. Among these components, CO2 and N2 are ubiquitous on earth. Thus, SCU_iGEM team aim to create a pioneer organism which is able to carry out carbon fixation and nitrogen fixation simultaneously. After inorganic carbon and nitrogen are fixed into organic substances, they could act as nutrients server for other immigrant organisms in the future.</p> | + | <p>The mean function of carbon fixation E.pangu is transforming inorganic substances into organic substances. Unfortunately, the process will definitely be affected by their natural ability of utilizing glucose to serve energy to E.pangu themselves. Thus, our project includes a part in which we will knockout the genes of glycolysis to verify the functions of carbon fixation E.pangu. The gene circuit for this part is shown in figure 1:</p> |
| + | <img src="https://static.igem.org/mediawiki/2015/b/ba/7BT-135SP0BQF%60-A8-~VY-5.png" title = "Fig. 1. Gene circuit of carbon fixation testing" /> |
| + | <h6 class="back"><strong>Fig. 1. Gene circuit of carbon fixation testing</strong></h6> |
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| + | <p>Here we introduced a gene editing technique called CRISPR Cas9 to knock out glycolysis of E.pangu. CRISPR is widely used to edit genome based on the complexity of sgRNA and Cas9 protein. Unlike other gene edition technique, CRISPR can be designed in one dimension instead of stereoscopy. Thus, when targeting different sites, we just need to design corresponding sequences. After knocking out several key enzymes, we will be able to regulate the metabolism of E.pangu, so that the ability of carbon fixation of our engineered microbe can be qualified.</p> |
| + | <p>At the initial stage of bacterial growth, the energy requirement is relatively high. To ensure the normal growth at this stage, we added a regulated promoter before Cas9 protein, through which the glucose catabolism of our bacteria can be regulated. That is, we will not knock out the glycolysis system until the cell growth has reached stationary phase.</p> |
| <hr style="border-bottom: 1px solid #DA6426;margin-bottom: 60px;margin-top: 60px; margin-left:-35px;margin-right: -35px;"> | | <hr style="border-bottom: 1px solid #DA6426;margin-bottom: 60px;margin-top: 60px; margin-left:-35px;margin-right: -35px;"> |
− | <h2><strong>Project overview</strong></h2> | + | <h3 class="back"><strong>Results</strong></h3> |
| + | <h4 class="back"><strong> 1.pRhl Insight</strong></h4> |
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| <p>In this summer, we are going to design a system which is able to fix inorganic carbon and nitrogen simultaneously. This system consists of 2 types of E.coli: nitrogen fixation E.coli and carbon fixation E.coli. We hope this system will play the role of pioneer organism on Mars immigration. Based on this purpose, we call them E. pangu collectively, which is named after the creator of universe, Pangu in Chinese mythology, as they will reclaim Mars for future immigrated organisms. </p> | | <p>In this summer, we are going to design a system which is able to fix inorganic carbon and nitrogen simultaneously. This system consists of 2 types of E.coli: nitrogen fixation E.coli and carbon fixation E.coli. We hope this system will play the role of pioneer organism on Mars immigration. Based on this purpose, we call them E. pangu collectively, which is named after the creator of universe, Pangu in Chinese mythology, as they will reclaim Mars for future immigrated organisms. </p> |
| <h3><strong>Our project contains the following contents:</strong></h3> | | <h3><strong>Our project contains the following contents:</strong></h3> |