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| <b>Leaders:</b> Yixian Yang, Zhandong Jiao<br> | | <b>Leaders:</b> Yixian Yang, Zhandong Jiao<br> |
| <b>Members:</b> Celine Siu Ham Zhang, Jiangshan Gao, Sizhe Duan, Marco Xue, Yeqi Wong, Xinyuan Wang, Bert Shan, James Haorui Wang, Junfei (Sophie) Li, Yiming Tang, Shao Chang, Siyuan Wang, Bohan Zhang<br> | | <b>Members:</b> Celine Siu Ham Zhang, Jiangshan Gao, Sizhe Duan, Marco Xue, Yeqi Wong, Xinyuan Wang, Bert Shan, James Haorui Wang, Junfei (Sophie) Li, Yiming Tang, Shao Chang, Siyuan Wang, Bohan Zhang<br> |
− | <b>Primary PI:</b> Shiqi Wang.<br> | + | <b>Primary PI:</b> Shiqi Wang<br> |
| <b>Instructors:</b> Xing (Leaves) Zhang, Tianze Zhu<br> | | <b>Instructors:</b> Xing (Leaves) Zhang, Tianze Zhu<br> |
| <b>Advisors:</b> Zhimeng Xu, Jun Ma, Aiqi Zhao, Li TianHong, Luo Shaowei, Weizhao Chen<br><br> | | <b>Advisors:</b> Zhimeng Xu, Jun Ma, Aiqi Zhao, Li TianHong, Luo Shaowei, Weizhao Chen<br><br> |
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| </div> | | </div> |
| <div class="subsection two_thirds"> | | <div class="subsection two_thirds"> |
− | There are several bio-preservation methods for bacteria. Nevertheless, at research institutes, ultra-low temperature freezing is the most commonly used method for long-term storage. However, the storage relies on specialized equipments, including the ultralow temperature refrigerators (-70℃~-80℃). Ultralow temperature refrigerators are expensive, noisy, and consumes lots of electricity, so there is usually no such equipment outside of laboratories.<br> Though there are some biopreservation means besides ultralow temperature, each method has shortcomings. This promoted us to initiate our project.</div> | + | 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> |
| <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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| alt=""></div> | | alt=""></div> |
| <div class="subsection two_thirds"> | | <div class="subsection two_thirds"> |
− | The metabolism slows down if the cells lose water. Indeed, there are some dried bacteria products on the market. Thus, we decided to make engineered bacteria into dry powder. Then the powder can be stored at room temperature without any professional equipment. | + | The metabolism slows down if the cells lose water. Indeed, we can find some dried bacteria products on the market. Thus, we decided to make engineered bacteria into dry powder. Then the powder can be stored at room temperature without any professional equipment. |
| </div> | | </div> |
| <div class="clear"></div> | | <div class="clear"></div> |
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| <img src="https://static.igem.org/mediawiki/2020/f/f4/T--QHFZ-China--Poster-sec2_4.png" | | <img src="https://static.igem.org/mediawiki/2020/f/f4/T--QHFZ-China--Poster-sec2_4.png" |
| alt=""> | | alt=""> |
− | </div> | + | </div><br> |
− | <div class="subsection two_thirds"> | + | <div class="subsection two_thirds"><br> |
| The direct drying method is time-consuming, and the metabolism will not stop during the process. However, freeze-drying is very fast. Indeed, freeze-drying has been widely used to produce vaccines, medicaments, and food because it can maintain the materials' original state. | | The direct drying method is time-consuming, and the metabolism will not stop during the process. However, freeze-drying is very fast. Indeed, freeze-drying has been widely used to produce vaccines, medicaments, and food because it can maintain the materials' original state. |
| </div> | | </div> |
| <div class="clear"></div> | | <div class="clear"></div> |
− | Nonetheless, during the process, at least three stresses affect cells' survival: freeze, desiccation, and vacuum. Several studies have tried to improve the survival rate of bacteria. Firstly, researchers tried to improve the procedure of freeze-drying; Secondly, they tried to optimize the constituent of protectants and excipients in the solution before lyophilization, but the survival rate still stayed at about 10%. Thus, we hope to introduce another element to elaborate on previous studies.<br><br> | + | Nonetheless, during the process, at least three stresses affect cells' survival: freeze, desiccation and vacuum. Several studies have tried to improve the survival rate of bacteria: (1) to improve the procedure of freeze-drying; (2) to optimize the constituent of protectants and excipients in the solution before lyophilization. Yet, the survival rate still stayed at about 10%. Thus, we hope to introduce another element to elaborate on previous studies. <br><br> |
| | | |
| <b>Tardigrade (water bear) / TDPs</b><br><br> | | <b>Tardigrade (water bear) / TDPs</b><br><br> |
| <div class="subsection third"><img src="https://static.igem.org/mediawiki/2020/c/c6/T--QHFZ-China--Poster-sec2_6.png" | | <div class="subsection third"><img src="https://static.igem.org/mediawiki/2020/c/c6/T--QHFZ-China--Poster-sec2_6.png" |
| alt=""></div> | | alt=""></div> |
− | <div class="subsection two_thirds"> To apply lyophilization/freeze-drying, we need to utilize the proteins of a very special organism. Tardigrade (water bear) is a miraculous organism found in 1773 and is famous for its strong vitality. It can survive extreme environments, like freezing, desiccation, vacuuming, extreme temperatures, high pressure, and radiation.</div> | + | <div class="subsection two_thirds"> To apply lyophilization/freeze-drying, we need to utilize the proteins of a very special organism. Tardigrade (water bear) is a miraculous organism found in 1773. It can survive extreme environments, like freezing, desiccation, vacuuming, extreme temperatures, high pressure, radiation, <i>etc</i>.</div> |
| <div class="clear"></div> | | <div class="clear"></div> |
| The strong vitality of tardigrades at least partially owes to TDPs (Tardigrade intrinsically Disordered Proteins), which are exceptionally resistant to desiccation.<br> | | The strong vitality of tardigrades at least partially owes to TDPs (Tardigrade intrinsically Disordered Proteins), which are exceptionally resistant to desiccation.<br> |
| + | TDPs are found in 2017, mainly including several Cytosolic-abundant heat soluble proteins (CAHS) and secreted massive heat soluble proteins (SAHS). TDPs were proven to protect living cells from drying<sup>[1]</sup> and maintain the activeness of proteins after freeze-drying<sup>[2,3]</sup>, but no one studied whether they can protect living cells from freeze-drying or not. Therefore, we tried to allow bacteria to express TDPs and tested whether they can survive freeze-drying and the subsequent storage processes.<br> |
| <img src="https://static.igem.org/mediawiki/2020/d/dc/T--QHFZ-China--Poster-sec2_7.png" | | <img src="https://static.igem.org/mediawiki/2020/d/dc/T--QHFZ-China--Poster-sec2_7.png" |
| alt=""><br> | | alt=""><br> |
− | TDPs are found in 2017, mainly including several Cytosolic-abundant heat soluble proteins (CAHS) and secreted massive heat soluble proteins (SAHS). TDPs were proven to protect living cells from drying<sup>[1]</sup> and maintain the activeness of proteins after freeze-drying<sup>[2,3]</sup>, but no one studied whether they can protect living cells from freeze-drying or not. Therefore, we tried to allow bacteria to express TDPs and tested whether they can survive freeze-drying and the subsequent storage processes.<br> | + | In this project, we studied 5 TDPs: CAHS 89226, CAHS 94205, CAHS 106094, CAHS 107838, and SAHS 33020, as well as two gene parts that are used before TDPs were found: LEA (Late Embryogenesis Abundant protein) from <i> Glycine max </i><sup>[4]</sup> and OtsBA (producing trehalose)<sup>[5]</sup>.<br> |
| <img src="https://static.igem.org/mediawiki/2020/2/21/T--QHFZ-China--Poster-sec2_8.png" | | <img src="https://static.igem.org/mediawiki/2020/2/21/T--QHFZ-China--Poster-sec2_8.png" |
− | alt=""><br>
| + | alt=""><br> |
− | In this project, we studied 5 TDPs: CAHS 89226, CAHS 94205, CAHS 106094, CAHS 107838, and SAHS 33020, as well as two gene parts that are used before TDPs were found: LEA (Late Embryogenesis Abundant protein) from <i> Glycine max </i><sup>[4]</sup> and OtsBA (producing trehalose)<sup>[5]</sup>.<br><br>
| + | |
| </div> | | </div> |
| </div> | | </div> |
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| <b>Leaders:</b> Yixian Yang, Zhandong Jiao<br> | | <b>Leaders:</b> Yixian Yang, Zhandong Jiao<br> |
| <b>Members:</b> Celine Siu Ham Zhang, Jiangshan Gao, Sizhe Duan, Marco Xue, Yeqi Wong, Xinyuan Wang, Bert Shan, James Haorui Wang, Junfei (Sophie) Li, Yiming Tang, Shao Chang, Siyuan Wang, Bohan Zhang<br> | | <b>Members:</b> Celine Siu Ham Zhang, Jiangshan Gao, Sizhe Duan, Marco Xue, Yeqi Wong, Xinyuan Wang, Bert Shan, James Haorui Wang, Junfei (Sophie) Li, Yiming Tang, Shao Chang, Siyuan Wang, Bohan Zhang<br> |
− | <b>Primary PI:</b> Shiqi Wang.<br> | + | <b>Primary PI:</b> Shiqi Wang<br> |
| <b>Instructors:</b> Xing (Leaves) Zhang, Tianze Zhu<br> | | <b>Instructors:</b> Xing (Leaves) Zhang, Tianze Zhu<br> |
| <b>Advisors:</b> Zhimeng Xu, Jun Ma, Aiqi Zhao, Li TianHong, Luo Shaowei, Weizhao Chen<br><br> | | <b>Advisors:</b> Zhimeng Xu, Jun Ma, Aiqi Zhao, Li TianHong, Luo Shaowei, Weizhao Chen<br><br> |
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| alt=""><br> | | alt=""><br> |
| <b>Basic model</b><br><br> | | <b>Basic model</b><br><br> |
− | With reasonable assumptions, we constructed a basic model to simulate the process. | + | With reasonable assumptions, we constructed a basic model to simulate the process. <br> |
| If the left part is activated, TDP (CAHS 106094) is expressed and finally reaches the equilibrium. We can change the strengths of Pc and RBS 1 to achieve different TDP levels<sup>[7,8]</sup>.<br> | | If the left part is activated, TDP (CAHS 106094) is expressed and finally reaches the equilibrium. We can change the strengths of Pc and RBS 1 to achieve different TDP levels<sup>[7,8]</sup>.<br> |
| <img src="https://static.igem.org/mediawiki/2020/4/42/T--QHFZ-China--Poster-sec7_21.png" | | <img src="https://static.igem.org/mediawiki/2020/4/42/T--QHFZ-China--Poster-sec7_21.png" |
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| <img src="https://static.igem.org/mediawiki/2020/e/e6/T--QHFZ-China--Poster-sec9_24.png" | | <img src="https://static.igem.org/mediawiki/2020/e/e6/T--QHFZ-China--Poster-sec9_24.png" |
| alt=""><br> | | alt=""><br> |
− | (3) There are some dry microbiological products, such as dry yeast for fermentation, probiotics tablets for health and nitrobacteria capsule for fish culture. The method can enhance the survival rate of the microorganisms to reduce the production costs.<br> | + | <div class="subsection third"> |
− | <img src="https://static.igem.org/mediawiki/2020/9/9b/T--QHFZ-China--Poster-sec9_25.png" alt=""><br> | + | <img src="https://static.igem.org/mediawiki/2020/9/9b/T--QHFZ-China--Poster-sec9_25.png" alt=""> |
− | (4) There is a lack of cold chains since they are expensive and easy to malfunction. Our method can allow the transportation of engineered bacteria without the usage of cold chains so that transportation is easier. The engineered bacteria can be used more widely around the world.<br> | + | </div> |
− | <img src="https://static.igem.org/mediawiki/2020/f/f9/T--QHFZ-China--Poster-sec9_26.png" | + | <div class="subsection two_thirds"> |
− | alt=""><br>
| + | (3) There are some dry microbiological products, such as dry yeast for fermentation, probiotics tablets for health and nitrobacteria capsule for fish culture. The method can enhance the survival rate of the microorganisms to reduce the production costs. |
| + | </div> |
| + | <div class="clear"></div> |
| + | <div class="subsection third"> |
| + | <img src="https://static.igem.org/mediawiki/2020/f/f9/T--QHFZ-China--Poster-sec9_26.png" |
| + | alt=""> |
| + | </div> |
| + | <div class="subsection two_thirds"> |
| + | (4) There is a lack of cold chains since they are expensive and easy to malfunction. Our method can allow the transportation of engineered bacteria without the usage of cold chains so that transportation is easier. The engineered bacteria can be used more widely around the world. Meanwhile, NASA has an imagination to transport some organisms to the other planets. Our method would give help. |
| + | </div> |
| + | <div class="clear"></div> |
| </div> | | </div> |
| </div> | | </div> |