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| <h4>Sample without Dengue Virions:</h4> | | <h4>Sample without Dengue Virions:</h4> |
− | The AuNPs will not accumulate on the test line, and no color change will be found. The AuNPs will aggregate on control line via the interaction of PTRS-1 and E protein, so the control line turns red. | + | The AuNPs will not accumulate on the test line, and no color change will be found. The AuNPs will aggregate on the control line via the interaction of PTRS-1 and E protein, so the control line turns red. |
| <br><br> | | <br><br> |
| <h4>Sample with Dengue Virions:</h4> | | <h4>Sample with Dengue Virions:</h4> |
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| <img src="https://static.igem.org/mediawiki/2020/f/f2/T--CCU_Taiwan--Poster_Results1.png"> | | <img src="https://static.igem.org/mediawiki/2020/f/f2/T--CCU_Taiwan--Poster_Results1.png"> |
| <br><br> | | <br><br> |
− | The TR-PCR products were taken to AD-PCR. The AD-PCR product of TRS-110*3 TRS-151*7 with sizes ranging from 200-500 bp were extracted and ligated to TA vectors. | + | The TR-PCR products were taken to AD-PCR. The AD-PCR products of TRS-110*3 and TRS-151*7 with sizes ranging from 200-500 bp were extracted and ligated to TA vectors. |
| <br> | | <br> |
| <img src="https://static.igem.org/mediawiki/2020/6/6b/T--CCU_Taiwan--Poster_Results2.png"> | | <img src="https://static.igem.org/mediawiki/2020/6/6b/T--CCU_Taiwan--Poster_Results2.png"> |
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| We expressed E protein to use it in the control line. | | We expressed E protein to use it in the control line. |
| <br> | | <br> |
− | The weak bands from 63 to 48 kDa indicate the expression of the E protein with the HA tag, which has a size of 61.2 kDa. We further confirmed using Western blot with the anti-His tag antibodies. | + | The weak bands from 63 to 48 kDa indicate the E protein expression with the HA tag, which has a size of 61.2 kDa. We further confirmed using Western blot with the anti-His tag antibodies. |
| <br> | | <br> |
| <img src="https://static.igem.org/mediawiki/2020/f/f3/T--CCU_Taiwan--Poster_Results3.png"> | | <img src="https://static.igem.org/mediawiki/2020/f/f3/T--CCU_Taiwan--Poster_Results3.png"> |
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| In addition to the required E protein used in our kit, we attempted to express CLEC5A as a model protein to interact with E protein. | | In addition to the required E protein used in our kit, we attempted to express CLEC5A as a model protein to interact with E protein. |
| <br> | | <br> |
− | The strong bands on 26.6 kDa indicate expression of the CLEC5A extracellular domain with the Myc tag, which is also confirmed using western blot with the anti-His tag antibodies. | + | The strong bands on 26.6 kDa indicate the CLEC5A extracellular domain with the Myc tag, which is also confirmed using western blot with the anti-His tag antibodies. |
| <br> | | <br> |
| <img src="https://static.igem.org/mediawiki/2020/0/0c/T--CCU_Taiwan--Poster_Results4.png"> | | <img src="https://static.igem.org/mediawiki/2020/0/0c/T--CCU_Taiwan--Poster_Results4.png"> |
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| As a proof of concept that we are able to form the covalent bonds between the primary amines (from the PTRS) and AuNPs, we tried to conjugate the DNA primers with the modified AuNPs. We found that with the DNA conjugation, the Raman signals have a significant decrease. Although we have no model to explain this effect, we believe it resulted from interactions with the DNA, suggesting DNA can bind to AuNPs. | | As a proof of concept that we are able to form the covalent bonds between the primary amines (from the PTRS) and AuNPs, we tried to conjugate the DNA primers with the modified AuNPs. We found that with the DNA conjugation, the Raman signals have a significant decrease. Although we have no model to explain this effect, we believe it resulted from interactions with the DNA, suggesting DNA can bind to AuNPs. |
| <br><br> | | <br><br> |
− | We expressed green fluorescent protein (GFP) as a mock E protein to show that after modification the glass fiber membranes can bind to the primary amines (side-chain amines) from a peptide or protein. The intensities of emission correlate to the concentration of GFP using in the reactions, suggesting the conjugation experiments were successful. | + | We expressed green fluorescent protein (GFP) as a mock E protein to show that the modified glass fiber membranes can bind to the primary amines (sidechain amines) from a peptide or protein after modification. The intensities of emission correlate to GFP concentration using in the reactions, suggesting the conjugation experiments were successful. |
| <br> | | <br> |
| <img src="https://static.igem.org/mediawiki/2020/2/2a/T--CCU_Taiwan--Poster_Results5.png"> | | <img src="https://static.igem.org/mediawiki/2020/2/2a/T--CCU_Taiwan--Poster_Results5.png"> |
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| <div class="text"> | | <div class="text"> |
| <h4>Interviews</h4> | | <h4>Interviews</h4> |
− | We wanted to shape our project from different perspectives to achieve socially responsible research. Thus, we consulted with government officials, experts, and academic specialists to consider the application of our project in society. From the interviews, we understood the process and the standards for verification for medical devices from the Taiwan Food and Drug Administration (TFDA). Moreover, we learned about pretreatment of blood sample from The Dengue Prevention and Control Center. We also comprehended the modification of gold nanoparticles after consulting with experts in the Department of Chemistry and Biochemistry at National Chung Cheng University. | + | We wanted to shape our project from different perspectives to achieve socially responsible research. Thus, we consulted with government officials, experts, and academic specialists to consider the application of our project in society. From the interviews, we understood the process and the standards for verification for medical devices from the Taiwan Food and Drug Administration. Moreover, we learned about the pretreatment of the blood sample from The Dengue Prevention and Control Center. We also comprehended the modification of gold nanoparticles after consulting with experts in the Department of Chemistry and Biochemistry at National Chung Cheng University. |
| <br><br> | | <br><br> |
| <h4>Public Survey</h4> | | <h4>Public Survey</h4> |
− | We want to find out how much the public knows about dengue and their demands for a dengue virus detection kit. Thus, we created a questionnaire and received 248 questionnaires by sharing the it online and distributing it in the community. | + | We want to find out how much the public knows about dengue and their demands for a dengue virus detection kit. Thus, we created a questionnaire and received 248 questionnaires by sharing it online and distributing it in the community. |
| <br><br> | | <br><br> |
− | The results showed that nearly 70% of people did not know about severe dengue and nearly 70% did not know what kind of treatment is required for dengue fever. This shows that the public in Taiwan lack a common understanding of dengue, which leads to low awareness about the severity of this disease in Taiwan. This urged us to conduct outreach and education programs to popularize knowledge of dengue fever. | + | The results showed that nearly 70% of people did not know about severe dengue, and nearly 70% did not know what kind of treatment is required for dengue fever. This shows that the public in Taiwan lacks a common understanding of dengue, which leads to low awareness about the severity of this disease in Taiwan. This urged us to conduct outreach and education programs to popularize knowledge of dengue fever. |
| <br><br> | | <br><br> |
| From the survey, we also determined that our detection kit meets their needs, as it should have high sensitivity during the first five days of symptoms, which is when people would consult a doctor. | | From the survey, we also determined that our detection kit meets their needs, as it should have high sensitivity during the first five days of symptoms, which is when people would consult a doctor. |
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| <img src="https://static.igem.org/mediawiki/2020/1/1b/T--CCU_Taiwan--Poster_HP-6.jpeg"> | | <img src="https://static.igem.org/mediawiki/2020/1/1b/T--CCU_Taiwan--Poster_HP-6.jpeg"> |
| </div> | | </div> |
− | Taiwan is considered an aged society, as the aged population reached 16% in 2018. We invited the elderly in our community help us record a dengue fever information audio in Taiwanese Hokkien, targeting senior citizens, who mostly use this local dialect. | + | Taiwan is considered an ageing society, as the aged population reached 16% in 2018. We invited the elderly in our community to help us record a dengue fever information audio in Taiwanese Hokkien, targeting senior citizens, who mostly use this local dialect. |
| <br><br> | | <br><br> |
| <li><b>Children’s Storybook</b></li> | | <li><b>Children’s Storybook</b></li> |
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| <img src="https://static.igem.org/mediawiki/2020/4/43/T--CCU_Taiwan--Poster_HP-7.jpeg"> | | <img src="https://static.igem.org/mediawiki/2020/4/43/T--CCU_Taiwan--Poster_HP-7.jpeg"> |
| </div> | | </div> |
− | We created a children’s storybook to spark children’s interest in science. To reach to more children and increase the impact of our storybook, we translated it into English, Chinese, and Hindi. | + | We created a children’s storybook to spark children’s interest in science. To reach more children and increase the impact of our storybook, we translated it into English, Chinese, and Hindi. |
| <br><br> | | <br><br> |
| <li><b>Educational Activity</b></li> | | <li><b>Educational Activity</b></li> |
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| <br><br> | | <br><br> |
| <li><b>Radio Interview</b></li> | | <li><b>Radio Interview</b></li> |
− | We were invited by National Education Radio to introduce synthetic biology, iGEM competition and also our project. | + | We were invited by National Education Radio to introduce synthetic biology, iGEM competition, and also our project. |
| <br><br> | | <br><br> |
| <li><b>Communication</b></li> | | <li><b>Communication</b></li> |
− | We joined the 2020 Taiwan iGEM Conference, which provided us a chance to share our project and received lots of valuable advice from other teams and professors. We also participated in a panel discussion focusing on sustainable development goals (SDGs). This leads us to have a deeper understanding of SDGs. | + | We joined the 2020 Taiwan iGEM Conference, which provided us a chance to share our project, and received lots of valuable advice from other teams and professors. We also participated in a panel discussion focusing on sustainable development goals (SDGs). This leads us to have a deeper understanding of SDGs. |
| </ul> | | </ul> |
| </div> | | </div> |
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| <img src="https://static.igem.org/mediawiki/2020/4/47/T--CCU_Taiwan--Poster_Inclusion-1.jpg"> | | <img src="https://static.igem.org/mediawiki/2020/4/47/T--CCU_Taiwan--Poster_Inclusion-1.jpg"> |
| </div> | | </div> |
− | We wanted to reach out to communities that are underrepresented in science to allow them to participate in science. Thus, we went to a juvenile correction school to popularize science, as juvenile inmates lack opportunities to learn science. | + | We want to reach out to communities underrepresented in science to allow them to participate in science. Thus, we went to a juvenile correction school to popularize science, as juvenile inmates lack opportunities to learn science. |
| <br><br> | | <br><br> |
| From this activity, we found that inclusion in science communication is a two-way street. Science should include communities that lack access to science by eliminating the access barriers with diverse and engaging ways to bring science closer to them. In order to experience the beauty of science, underrepresented communities must overcome conventional thinking that science is too difficult. | | From this activity, we found that inclusion in science communication is a two-way street. Science should include communities that lack access to science by eliminating the access barriers with diverse and engaging ways to bring science closer to them. In order to experience the beauty of science, underrepresented communities must overcome conventional thinking that science is too difficult. |
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| <h4>Glove Puppetry</h4> | | <h4>Glove Puppetry</h4> |
| <img src="https://static.igem.org/mediawiki/2020/0/0c/T--CCU_Taiwan--Poster_Inclusion-2.jpeg"> | | <img src="https://static.igem.org/mediawiki/2020/0/0c/T--CCU_Taiwan--Poster_Inclusion-2.jpeg"> |
− | Glove puppetry is a traditional performing art in Taiwan with its own unique elements such as using the language most spoken by Taiwanese elderly, Taiwanese Hokkien, for narration. This inspired us to integrate this folk art into science communication. We thought that a non-scientific artist could also provide a different perspective on science. | + | Glove puppetry is a traditional performing art in Taiwan with its own unique elements, such as using the language most spoken by Taiwanese elderly, Taiwanese Hokkien, for narration. This inspired us to integrate this folk art into science communication. We thought that a non-scientific artist could also provide a different perspective on science. |
| <br><br> | | <br><br> |
| Thus, we collaborated with a glove puppetry troupe to transmit knowledge of dengue by narrating and filming a story as a 13-minute video. | | Thus, we collaborated with a glove puppetry troupe to transmit knowledge of dengue by narrating and filming a story as a 13-minute video. |
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| <li><b>Introduction</b></li> | | <li><b>Introduction</b></li> |
| <ol> | | <ol> |
− | <li>Taiwan National Infectious Disease Statistics System - Taiwan Centers of Disease Control (TCDC).</li>
| |
| <li>Dengue and severe dengue - World Health Organization.</li> | | <li>Dengue and severe dengue - World Health Organization.</li> |
− | <li>Dengue Fever – Taiwan Centers of Disease Control (TCDC).</li> | + | <li>Taiwan National Infectious Disease Statistics System - Taiwan Centers of Disease Control.</li> |
| + | <li>Dengue Fever – Taiwan Centers of Disease Control.</li> |
| </ol> | | </ol> |
| <br> | | <br> |
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| <ol> | | <ol> |
| <li>Szu-Ting Chen, Fei-Ju Li, Tzy-yun Hsu, Shu-Mei Liang, Yi-Chen Yeh, Wen-Yu Liao, Teh-Ying Chou, Nien-Jun Chen, Michael Hsiao, Wen-Bin Yang, and Shie-Liang Hsieh. CLEC5A is a critical receptor in innate immunity against Listeria infection. Nature Communications. 2017; 8(1): 299. doi:10.1038/s41467-017-00356-3.</li> | | <li>Szu-Ting Chen, Fei-Ju Li, Tzy-yun Hsu, Shu-Mei Liang, Yi-Chen Yeh, Wen-Yu Liao, Teh-Ying Chou, Nien-Jun Chen, Michael Hsiao, Wen-Bin Yang, and Shie-Liang Hsieh. CLEC5A is a critical receptor in innate immunity against Listeria infection. Nature Communications. 2017; 8(1): 299. doi:10.1038/s41467-017-00356-3.</li> |
− | <li>Aleksandra A Watson, Andrey A Lebedev, Benjamin A Hall, Angharad E Fenton-May, Alexei A Vagin, Wanwisa Dejnirattisai, James Felce, Juthathip Mongkolsapaya, Angelina S Palma, Yan Liu, Ten Feizi, Gavin R Screaton, Garib N Murshudov, and Christopher A O'Callaghan. Structural flexibility of the macrophage dengue virus receptor CLEC5A: implications for ligand binding and signaling. Journal of Biological Chemistry. 2011; 286(27): 24208-24218. doi: 10.1074/jbc.M111.226142.</li> | + | <li>Aleksandra A Watson, Andrey A Lebedev, Benjamin A Hall, Angharad E Fenton-May, Alexei A Vagin, Wanwisa Dejnirattisai, James Felce, Juthathip Mongkolsapaya, Angelina S Palma, Yan Liu, Ten Feizi, Gavin R Screaton, Garib N Murshudov, and Christopher A O'Callaghan. Structural flexibility of the macrophage dengue virus receptor CLEC5A: implications for ligand binding and signaling. Journal of Biological Chemistry. 2011; 286(27): 24208-24218. doi:10.1074/jbc.M111.226142.</li> |
| </ol> | | </ol> |
| <br> | | <br> |
| <li><b>LAE</b></li> | | <li><b>LAE</b></li> |
| <ol> | | <ol> |
− | <li>Peng-Yeh Lai, Chia-Tse Hsu, Shao-Hung Wang, Jin-Ching Lee, Min-Jen Tseng, Jaulang Hwang, Wen-Tsai Ji, and Hau-Ren Chen. Production of a neutralizing antibody against envelope protein of dengue virus type 2 using the linear array epitope technique. Journal of General Virology. 2014; 95(10): 2155-2165. doi: 10.1099/vir.0.062562-0.</li> | + | <li>Peng-Yeh Lai, Chia-Tse Hsu, Shao-Hung Wang, Jin-Ching Lee, Min-Jen Tseng, Jaulang Hwang, Wen-Tsai Ji, and Hau-Ren Chen. Production of a neutralizing antibody against envelope protein of dengue virus type 2 using the linear array epitope technique. Journal of General Virology. 2014; 95(10): 2155-2165. doi:10.1099/vir.0.062562-0.</li> |
| </ol> | | </ol> |
| <br><br> | | <br><br> |
| <li><b>Modeling: Interaction between the Tandem-repeated Sequence Peptide and E Protein</b></li> | | <li><b>Modeling: Interaction between the Tandem-repeated Sequence Peptide and E Protein</b></li> |
| <ol> | | <ol> |
− | <li>Maciej Ciemny, Mateusz Kurcinski, Karol Kamel, Andrzej Kolinski, Nawsad Alam, Ora Schueler-Furman, and Sebastian Kmiecik. Protein–peptide docking: opportunities and challenges. Drug Discovery Today. 2018; 23(8): 1530-1537. doi: 10.1016/j.drudis.2018.05.006.</li> | + | <li>Maciej Ciemny, Mateusz Kurcinski, Karol Kamel, Andrzej Kolinski, Nawsad Alam, Ora Schueler-Furman, and Sebastian Kmiecik. Protein–peptide docking: opportunities and challenges. Drug Discovery Today. 2018; 23(8): 1530-1537. doi:10.1016/j.drudis.2018.05.006.</li> |
− | <li>Steven A Combs, Samuel L Deluca, Stephanie H Deluca, Gordon H Lemmon, David P Nannemann, Elizabeth D Nguyen, Jordan R Willis, Jonathan H Sheehan, and Jens Meiler. Small-molecule ligand docking into comparative models with Rosetta. Nature Protocols. 2013; 8(7): 1277-1298. doi: 10.1038/nprot.2013.074.</li> | + | <li>Steven A Combs, Samuel L Deluca, Stephanie H Deluca, Gordon H Lemmon, David P Nannemann, Elizabeth D Nguyen, Jordan R Willis, Jonathan H Sheehan, and Jens Meiler. Small-molecule ligand docking into comparative models with Rosetta. Nature Protocols. 2013; 8(7): 1277-1298. doi:10.1038/nprot.2013.074.</li> |
| </ol> | | </ol> |
| <br> | | <br> |
| <li><b>Modeling: Interaction between Gold Nanoparticles</b></li> | | <li><b>Modeling: Interaction between Gold Nanoparticles</b></li> |
| <ol> | | <ol> |
− | <li>Elena Pokidysheva, Ying Zhang, Anthony J Battisti, Carol M Bator-Kelly, Paul R Chipman, Chuan Xiao, G Glenn Gregorio, Wayne A Hendrickson, Richard J Kuhn, and Michael G Rossmann. Cryo-EM Reconstruction of Dengue Virus in Complex with the Carbohydrate Recognition Domain of DC-SIGN. Cell. 2006; 124(3): 485-93. doi: 10.1016/j.cell.2005.11.042.</li> | + | <li>Elena Pokidysheva, Ying Zhang, Anthony J Battisti, Carol M Bator-Kelly, Paul R Chipman, Chuan Xiao, G Glenn Gregorio, Wayne A Hendrickson, Richard J Kuhn, and Michael G Rossmann. Cryo-EM Reconstruction of Dengue Virus in Complex with the Carbohydrate Recognition Domain of DC-SIGN. Cell. 2006; 124(3): 485-493. doi:10.1016/j.cell.2005.11.042.</li> |
| <li>Jörg Polte. Fundamental growth principles of colloidal metal nanoparticles – a new perspective. CrystEngComm. 2015, 17(36): 6809-6830. doi:10.1039/C5CE01014D.</li> | | <li>Jörg Polte. Fundamental growth principles of colloidal metal nanoparticles – a new perspective. CrystEngComm. 2015, 17(36): 6809-6830. doi:10.1039/C5CE01014D.</li> |
− | <li>Phillip E Mason, Adrien Lerbret, Marie-Louise Saboungi, George W Neilson, Christopher E Dempsey, and John W Brady. Glucose Interactions with a Model Peptide. Proteins. 2011; 79(7): 2224-2232. doi: 10.1002/prot.23047.</li> | + | <li>Phillip E Mason, Adrien Lerbret, Marie-Louise Saboungi, George W Neilson, Christopher E Dempsey, and John W Brady. Glucose Interactions with a Model Peptide. Proteins. 2011; 79(7): 2224-2232. doi:10.1002/prot.23047.</li> |
− | <li>Taehoon Kim, Kangtaek Lee, Myoung-seon Gong, and Sang-Woo Joo. Control of Gold Nanoparticle Aggregates by Manipulation of Interparticle Interaction. Langmuir . 2005; 21(21): 9524-9528. doi: 10.1021/la0504560.</li> | + | <li>Taehoon Kim, Kangtaek Lee, Myoung-seon Gong, and Sang-Woo Joo. Control of Gold Nanoparticle Aggregates by Manipulation of Interparticle Interaction. Langmuir. 2005; 21(21): 9524-9528. doi:10.1021/la0504560.</li> |
| </ol> | | </ol> |
| </ul> | | </ul> |