|
|
(3 intermediate revisions by the same user not shown) |
Line 311: |
Line 311: |
| 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"> |
Line 318: |
Line 318: |
| 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"> |
Line 327: |
Line 327: |
| 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 (sidechain 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"> |
Line 351: |
Line 351: |
| <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 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. | + | 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> |
Line 420: |
Line 420: |
| <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> |
Line 428: |
Line 428: |
| <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> |