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| <b>Abstract</b><br><br> | | <b>Abstract</b><br><br> |
− | Various engineered bacteria displayed useful functions. However, these bacteria's storage usually requires -80℃ refrigerators, which substantially limited their transportation and usages in daily life. Therefore, we decided to utilize TDPs from tardigrades (water bears) to supply a new storage method. We could produce dry life paused-bacteria powder by introducing the TDPs as protectants into bacteria during lyophilization (freeze-drying). The powder can then be stored at room temperature for a long time without any professional equipment usage.<br> | + | Various engineered bacteria displayed useful functions. However, these bacteria's storage usually requires -80℃ refrigerators, which substantially limited their transportation and usages in daily life. Therefore, we decided to utilize TDPs from tardigrades (water bears) to supply a new storage method. We could produce dry life-paused bacteria powder by introducing the TDPs as protectants into bacteria during lyophilization (freeze-drying). The powder can then be stored at room temperature for a long time without any professional equipment usage.<br> |
| This year, we confirmed that certain TDPs could maintain the survival rate of <i>E. coli</i>. We also optimized the method by regulating the expression levels and combining different TDPs. Also, we proved the modularity and tried to regulate the degradation of TDPs after freeze-drying. Considering that many iGEM teams and researchers are designing engineered bacteria for various application scenarios out of the laboratory, we hope our improvement in bio-preservation methods would promote their practical applications.<br><br> | | This year, we confirmed that certain TDPs could maintain the survival rate of <i>E. coli</i>. We also optimized the method by regulating the expression levels and combining different TDPs. Also, we proved the modularity and tried to regulate the degradation of TDPs after freeze-drying. Considering that many iGEM teams and researchers are designing engineered bacteria for various application scenarios out of the laboratory, we hope our improvement in bio-preservation methods would promote their practical applications.<br><br> |
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| <div class="title">Introduction & Inspiration</div> | | <div class="title">Introduction & Inspiration</div> |
| <div class="text"> | | <div class="text"> |
− | Our project is a continuation of the project created by QHFZ-2019. Last year, we made a portable household uric acid detector, which is a genetically engineered bacteria. It would facilitate the recovery of gout patients. However, we realized that there are still some problems hindering its practical application. Among them, we wanted to solve the issue of room-temperature storage before anything else.<br> | + | Our project is a continuation of the project created by QHFZ-2019. Last year, we made a portable household uric acid detector, which is a genetically engineered bacterium. It would facilitate the recovery of gout patients. However, we realized that there are still some problems hindering its practical application. Among them, we wanted to solve the issue of room-temperature storage before anything else.<br> |
| <img src="https://static.igem.org/mediawiki/2020/2/27/T--QHFZ-China--Poster_section1_1.png" | | <img src="https://static.igem.org/mediawiki/2020/2/27/T--QHFZ-China--Poster_section1_1.png" |
| alt=""><br> | | alt=""><br> |
− | Engineered bacteria need to be able to store for an extended period before being used at room temperature. They should stay alive while their metabolism paused. In laboratories, we use ultralow temperature to achieve this. That is why -80℃ refrigerator is necessary for molecular biology labs. However, it is hard to utilize such professional equipment from laboratories since they cannot be found daily. In general, household refrigerators can only achieve about -20℃, which is insufficient. Besides, it is not ideal to use the refrigerators to store bacteria, which we primarily use to store food. However, many engineered bacteria are designed for our daily life. We interviewed ten iGEM teams, and most of them believed that a suitable method to solve the problem would benefit the practical application of their products.<br> | + | Engineered bacteria need to be stored for an extended period before being used at room temperature. They should stay alive while their metabolism paused. In laboratories, we use ultralow temperature to achieve this. That is why -80℃ refrigerator is necessary for molecular biology labs. However, it is hard to utilize such professional equipment from laboratories since they cannot be found daily. In general, household refrigerators can only achieve about -20℃, which is insufficient. Besides, it is not ideal to use the refrigerators to store bacteria, which we primarily use to store food. However, many engineered bacteria are designed for our daily life. We interviewed ten iGEM teams, and most of them believed that a suitable method to solve the problem would benefit the practical application of their products.<br> |
| <img src="https://static.igem.org/mediawiki/2020/2/2b/T--QHFZ-China--Poster_section1_2.png" | | <img src="https://static.igem.org/mediawiki/2020/2/2b/T--QHFZ-China--Poster_section1_2.png" |
| alt=""><br> | | alt=""><br> |
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| </div> | | </div> |
| <div class="subsection two_thirds"> | | <div class="subsection two_thirds"> |
− | Ultra-low temperature freezing is the most commonly used long-term bio-preservation method at research institutes. However, the storage relies on specialized equipments, including the ultralow temperature refrigerators, but there is usually no such equipment outside of laboratories.<br> Though there are some other biopreservation means, each method has its own shortcomings.</div> | + | Ultra-low temperature freezing is the most commonly used long-term bio-preservation method at research institutes. However, the storage relies on specialized equipment, including the ultralow temperature refrigerators, but there is usually no such equipment outside of laboratories.<br> Though there are some other biopreservation means, each method has its own shortcomings.</div> |
| <div class="clear"></div> | | <div class="clear"></div> |
| <b>Freeze-drying / Lyophilization</b><br><br> | | <b>Freeze-drying / Lyophilization</b><br><br> |
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| <b>Abstract</b><br><br> | | <b>Abstract</b><br><br> |
− | Various engineered bacteria displayed useful functions. However, these bacteria's storage usually requires -80℃ refrigerators, which substantially limited their transportation and usages in daily life. Therefore, we decided to utilize TDPs from tardigrades (water bears) to supply a new storage method. We could produce dry life paused-bacteria powder by introducing the TDPs as protectants into bacteria during lyophilization (freeze-drying). The powder can then be stored at room temperature for a long time without any professional equipment usage.<br> | + | Various engineered bacteria displayed useful functions. However, these bacteria's storage usually requires -80℃ refrigerators, which substantially limited their transportation and usages in daily life. Therefore, we decided to utilize TDPs from tardigrades (water bears) to supply a new storage method. We could produce dry life-paused bacteria powder by introducing the TDPs as protectants into bacteria during lyophilization (freeze-drying). The powder can then be stored at room temperature for a long time without any professional equipment usage.<br> |
| This year, we confirmed that certain TDPs could maintain the survival rate of <i>E. coli</i>. We also optimized the method by regulating the expression levels and combining different TDPs. Also, we proved the modularity and tried to regulate the degradation of TDPs after freeze-drying. Considering that many iGEM teams and researchers are designing engineered bacteria for various application scenarios out of the laboratory, we hope our improvement in bio-preservation methods would promote their practical applications.<br><br> | | This year, we confirmed that certain TDPs could maintain the survival rate of <i>E. coli</i>. We also optimized the method by regulating the expression levels and combining different TDPs. Also, we proved the modularity and tried to regulate the degradation of TDPs after freeze-drying. Considering that many iGEM teams and researchers are designing engineered bacteria for various application scenarios out of the laboratory, we hope our improvement in bio-preservation methods would promote their practical applications.<br><br> |
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| <div class="text"> | | <div class="text"> |
| <b>Acknowledgement</b><br><br> | | <b>Acknowledgement</b><br><br> |
| + | To accomplish the project, we did most of the work. However, without others help, we cannot reach this achievement.<br> |
| We thank<br> | | We thank<br> |
− | Our teachers: Shiqi Wang, Xing (Leaves) Zhang, Tianze Zhu, Zhimeng Xu, Jun Ma, Aiqi Zhao, Li | + | <b>Our teachers:</b> Shiqi Wang, Xing (Leaves) Zhang, Tianze Zhu, Zhimeng Xu, Jun Ma, Aiqi Zhao, Li |
| TianHong, Luo Shaowei and Weizhao Chen<br> | | TianHong, Luo Shaowei and Weizhao Chen<br> |
− | Our interviewees: Guangyuna Song, Jiajia Liu, Miatta Momoh, Shan Jiang, Shaojie Li, Tianqi | + | <b>Our interviewees:</b> Guangyuna Song, Jiajia Liu, Miatta Momoh, Shan Jiang, Shaojie Li, Tianqi |
− | Zhang, Yikai Bao, ZhiZhong Gong;<br>Cooperating teams: Jilin_China, ZJUT_China_B, NEFU-China, | + | Zhang, Yikai Bao, ZhiZhong Gong;<br> |
| + | <b>Cooperating teams:</b> Jilin_China, ZJUT_China_B, NEFU-China, |
| XMU-China, ASTWS-China, BNDS_China, NAU-China, SHSBNU_China, SCU-China, NEU_China, | | XMU-China, ASTWS-China, BNDS_China, NAU-China, SHSBNU_China, SCU-China, NEU_China, |
| ECUST_China, KEYSTONE, BHSF, BJ101HS, NFLS, Tsinghua, BUCT-China, JNFLS, Nanjing_NFLS, | | ECUST_China, KEYSTONE, BHSF, BJ101HS, NFLS, Tsinghua, BUCT-China, JNFLS, Nanjing_NFLS, |
| RDFZ-China, QHFZ-China (2019), BNU-China (2019) and BEAS_China (2019)<br> | | RDFZ-China, QHFZ-China (2019), BNU-China (2019) and BEAS_China (2019)<br> |
− | The organizations helping us: iGEM Foundation, Tsinghua University High School, ZENO | + | <b>The organizations helping us:</b> iGEM Foundation, Tsinghua University High School, ZENO |
| CO.,LTD, BEIJING TIDE PHARMACEUTICAL CO.,LTD, YOUCARE PHARMACEUTICAL GROUP CO.,LTD, | | CO.,LTD, BEIJING TIDE PHARMACEUTICAL CO.,LTD, YOUCARE PHARMACEUTICAL GROUP CO.,LTD, |
| MathsWorks, Snapgene, Geneious, Beijing Haidian vocational school, Beijing Bayi School | | MathsWorks, Snapgene, Geneious, Beijing Haidian vocational school, Beijing Bayi School |
| Affiliated Yuquan Middle School, Tsinghua University<br> | | Affiliated Yuquan Middle School, Tsinghua University<br> |
− | and all people who gave us help.<br> | + | <b>and</b> all people who gave us help.<br> |
| Please accept our deepest thanks to everyone.<br><br> | | Please accept our deepest thanks to everyone.<br><br> |
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