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| <img src="https://static.igem.org/mediawiki/2020/9/99/T--CCU_Taiwan--Poster_Introduction.png"> | | <img src="https://static.igem.org/mediawiki/2020/9/99/T--CCU_Taiwan--Poster_Introduction.png"> |
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− | Dengue fever can progress into more serious conditions like dengue hemorrhagic fever (DHF), which has a 20% mortality rate. If conditions continue to worsen, patients experience dengue shock syndrome (DSS), which has a mortality rate of up to 40%. An estimated 500,000 patients develop DHF or DSS, resulting in about 25,000 deaths every year. | + | Dengue fever can progress into more severe conditions like dengue hemorrhagic fever (DHF), which has a 20% mortality rate. If conditions continue to worsen, patients experience dengue shock syndrome (DSS), which has a mortality rate of up to 40%. An estimated 500,000 patients develop DHF or DSS, resulting in about 25,000 deaths every year. |
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| Taiwan is in the area under the threat of the dengue virus. From 2014 to 2015, there were over 44,000 confirmed cases during a dengue fever outbreak that spread from Tainan to other cities, causing 228 deaths. Recently, there was a local transmission of dengue fever in Taoyuan, showing that the potentially affected population keeps increasing. | | Taiwan is in the area under the threat of the dengue virus. From 2014 to 2015, there were over 44,000 confirmed cases during a dengue fever outbreak that spread from Tainan to other cities, causing 228 deaths. Recently, there was a local transmission of dengue fever in Taoyuan, showing that the potentially affected population keeps increasing. |
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| After infection by the dengue virus, a nonstructural protein, NS1, appears in blood serum early in the infection. The current method of detecting dengue fever uses antibodies to recognize NS1 antigens. Nevertheless, the cost of the antibodies is high, the antibodies are hard to purify, and the procedure of making antibodies is time-consuming. | | After infection by the dengue virus, a nonstructural protein, NS1, appears in blood serum early in the infection. The current method of detecting dengue fever uses antibodies to recognize NS1 antigens. Nevertheless, the cost of the antibodies is high, the antibodies are hard to purify, and the procedure of making antibodies is time-consuming. |
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− | We took a step back to seek an alternative. We aimed to find something smaller and easier to produce to detect the dengue virus instead of using antibodies. Symptoms of DHF and DSS result from the interaction between C-type lectin domain, family 5, member A (CLEC5A) on macrophages and the envelope protein (E proteins) of dengue virus. | + | We took a step back to seek an alternative. We aimed to find something smaller and easier to produce to detect the dengue virus instead of antibodies. Symptoms of DHF and DSS result from the interaction between C-type lectin domain, family 5, member A (CLEC5A) on macrophages and the envelope protein (E proteins) of dengue virus. |
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| <div id="imginfo"> | | <div id="imginfo"> |
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| </div> | | </div> |
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− | We found the sequences from CLEC5A most likely to bind the dengue virus and confirmed this using Rosetta simulations. We believe these peptides have potential to detect the dengue virus. | + | We found the sequences from CLEC5A most likely to bind the dengue virus and confirmed these using Rosetta simulations. We believe these peptides have the potential to detect the dengue virus. |
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− | We designed a detection kit with these peptides that can bind to the E protein of dengue virus. This would allow people who may have dengue fever or live in an outbreak region of dengue fever to get treatment or protection and control the spread early. | + | We designed a detection kit with these peptides that can bind to the E protein of the dengue virus. This would allow people who may have dengue fever or live in an outbreak region of dengue fever to get treatment or protection and control the spread early. |
| </div> | | </div> |
| </div> | | </div> |
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| </ol> | | </ol> |
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− | For the primer design, TR-PCR primers need to be partially complementary to each other and AD-PCR primers need to have restriction sites. | + | For the primer design, TR-PCR primers need to be partially complementary to each other, and AD-PCR primers need to have restriction sites. |
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| <img src="https://static.igem.org/mediawiki/2020/2/2d/T--CCU_Taiwan--Poster_LAE-1.gif"> | | <img src="https://static.igem.org/mediawiki/2020/2/2d/T--CCU_Taiwan--Poster_LAE-1.gif"> |
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− | In TR-PCR, those primers act as both primer and template and they are partially complementary. The TRSs can be produced during the PCR cycling process. In AD-PCR, restriction sites need be introduced into the products of TR-PCR by adding the AD-PCR primers in PCR. | + | In TR-PCR, those primers act as both primer and template, and they are partially complementary. The TRSs can be produced during the PCR cycling process. In AD-PCR, restriction sites need be be introduced into the products of TR-PCR by adding the AD-PCR primers in PCR. |
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| <img src="https://static.igem.org/mediawiki/2020/4/4c/T--CCU_Taiwan--Poster_LAE-2.gif"> | | <img src="https://static.igem.org/mediawiki/2020/4/4c/T--CCU_Taiwan--Poster_LAE-2.gif"> |
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| <div class="title">BioBricks</div> | | <div class="title">BioBricks</div> |
| <div class="text"> | | <div class="text"> |
− | Our peptides and proteins were expressed in <i>E. Coli</i> with T7 promotor and lac operator, which can be induced by IPTG. The thrombin site was included to remove unnecessary sections. The His tag was used in Western blot to confirm the expression of protein, and the HA tag and myc tag were utilized for purification. | + | Our peptides and proteins were expressed in <i>E. Coli</i> with a T7 promoter and a lac operator, which can be induced by IPTG. The thrombin site was included to remove unnecessary sections. The His tag was used in Western blot to confirm the expression of the protein, and the HA tag and myc tag were utilized for purification. |
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| <img src="https://static.igem.org/mediawiki/2020/8/85/T--CCU_Taiwan--Poster_Parts-1.png"> | | <img src="https://static.igem.org/mediawiki/2020/8/85/T--CCU_Taiwan--Poster_Parts-1.png"> |