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| <div class="text"><p style="font-family:roboto;font-size:18px;"><span style="color:#006400"><b>Anti-Microbial Resistance (AMR): A Global Concern</b></span><br><br></p><div class="child"><img src="https://static.igem.org/mediawiki/2020/a/ad/T--IISER-Tirupati_India--Poster_motivation1.png" align="top"> | | <div class="text"><p style="font-family:roboto;font-size:18px;"><span style="color:#006400"><b>Anti-Microbial Resistance (AMR): A Global Concern</b></span><br><br></p><div class="child"><img src="https://static.igem.org/mediawiki/2020/a/ad/T--IISER-Tirupati_India--Poster_motivation1.png" align="top"> |
| </div><div class="clear"></div><br> | | </div><div class="clear"></div><br> |
− | <p style="font-family:roboto;font-size:16px;"><span style="color:#006400"><b>Fig 1:</b> Deaths attributable to AMR every year compared to other major causes of death<sup>4</sup>. | + | <p style="font-family:roboto;font-size:16px;"><span style="color:#006400"><b>Fig :</b> Deaths attributable to AMR every year compared to other major causes of death<sup>4</sup>. |
| <br>Antimicrobials help treat and prevent serious infections. As bacteria that cause infection adapt to withstand antibiotics, the potential for antimicrobial resistance to cause a global health crisis looms large.</span><br><br> | | <br>Antimicrobials help treat and prevent serious infections. As bacteria that cause infection adapt to withstand antibiotics, the potential for antimicrobial resistance to cause a global health crisis looms large.</span><br><br> |
| With the current of COVID-19 pandemic situation, there is a huge potential that certain activities could drive antimicrobial resistance even further<sup>1</sup>. In addition, epigenetic modifications<sup>2</sup>and biofilm formation<sup>3</sup> further increase bacterial resistance . Phage therapy which was thought to be the best solution is also failing due to CRISPR and other innate bacterial defenses<sup>4</sup>. | | With the current of COVID-19 pandemic situation, there is a huge potential that certain activities could drive antimicrobial resistance even further<sup>1</sup>. In addition, epigenetic modifications<sup>2</sup>and biofilm formation<sup>3</sup> further increase bacterial resistance . Phage therapy which was thought to be the best solution is also failing due to CRISPR and other innate bacterial defenses<sup>4</sup>. |
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| <div class="subsection full"><img src=" https://static.igem.org/mediawiki/2020/d/d1/T--IISER-Tirupati_India--Poster_motivation2.png" align="top"> | | <div class="subsection full"><img src=" https://static.igem.org/mediawiki/2020/d/d1/T--IISER-Tirupati_India--Poster_motivation2.png" align="top"> |
| </div> | | </div> |
− | <p style="font-family:roboto;font-size:16px;"><span style="color:#006400"><b>Fig 2:</b> Number of formulation companies manufacturing various antibiotics for animal use. Sulfonamides being one the highly produced antibiotics in India<sup>5</sup>.</span><br> | + | <p style="font-family:roboto;font-size:16px;"><span style="color:#006400"><b>Fig :</b> Number of formulation companies manufacturing various antibiotics for animal use. Sulfonamides being one the highly produced antibiotics in India<sup>5</sup>.</span><br> |
| <br>AMR emergence in India is majorly driven by factors like the high burden of bacterial infections, poor sanitary, hygiene conditions, and the increasing proportion of intensive animal farming, and this is ramped by the unregulated access to antibiotics, including sale without prescription or with an invalid prescription.<br><br> | | <br>AMR emergence in India is majorly driven by factors like the high burden of bacterial infections, poor sanitary, hygiene conditions, and the increasing proportion of intensive animal farming, and this is ramped by the unregulated access to antibiotics, including sale without prescription or with an invalid prescription.<br><br> |
| <b><span style="font-family:roboto;font-size:18px;color:#006400;">AMR & Livestock Farms</span></b><br></p> | | <b><span style="font-family:roboto;font-size:18px;color:#006400;">AMR & Livestock Farms</span></b><br></p> |
| <div class="child"><img src=" https://static.igem.org/mediawiki/2020/7/78/T--IISER-Tirupati_India--Poster_motivation3.png" align="top"> | | <div class="child"><img src=" https://static.igem.org/mediawiki/2020/7/78/T--IISER-Tirupati_India--Poster_motivation3.png" align="top"> |
| </div><div class="clear"></div> | | </div><div class="clear"></div> |
− | <p style="font-family:roboto;font-size:16px;"><span style="color:white"><b>Fig 3:</b> More than 73% of all antimicrobials sold in the world are used in animals<sup>6</sup><br>.</span><span style="color:#006400"><b>Fig 3:</b> More than 73% of all antimicrobials sold in the world are used in animals<sup>6</sup>.</span><br> | + | <p style="font-family:roboto;font-size:16px;"><span style="color:white"><b>Fig :</b> More than 73% of all antimicrobials sold in the world are used in animals<sup>6</sup><br>.</span><span style="color:#006400"><b>Fig :</b> More than 73% of all antimicrobials sold in the world are used in animals<sup>6</sup>.</span><br> |
| <br>The use of antibiotics as growth promoters in animal husbandry is a common practice and this, in turn, affects human health, as antibiotic-resistant bacteria can be transmitted between humans and animals through contact, food products, and from the environment.<sup>10</sup><br><br> | | <br>The use of antibiotics as growth promoters in animal husbandry is a common practice and this, in turn, affects human health, as antibiotic-resistant bacteria can be transmitted between humans and animals through contact, food products, and from the environment.<sup>10</sup><br><br> |
| <b> <span style="font-family:roboto;font-size:18px;color:#006400;">Antibiotics in Agricultural Waste</span></b><br></p> | | <b> <span style="font-family:roboto;font-size:18px;color:#006400;">Antibiotics in Agricultural Waste</span></b><br></p> |
| <div class="subsection full"><img src=" https://static.igem.org/mediawiki/2020/8/83/T--IISER-Tirupati_India--Poster_motivation4.png" align="top"> | | <div class="subsection full"><img src=" https://static.igem.org/mediawiki/2020/8/83/T--IISER-Tirupati_India--Poster_motivation4.png" align="top"> |
| </div> | | </div> |
− | <p style="font-family:roboto;font-size:16px;"><span style="color:#006400"><b>Fig 4:</b> The graph depicts the detection of antibiotics in poultry: Poultry Litter, swine: Swine Manure, and beef cattle: Cattle Manure manure<sup>8</sup>.</span> | + | <p style="font-family:roboto;font-size:16px;"><span style="color:#006400"><b>Fig :</b> The graph depicts the detection of antibiotics in poultry: Poultry Litter, swine: Swine Manure, and beef cattle: Cattle Manure manure<sup>8</sup>.</span> |
| <br> <span style="font-family:roboto;font-size:14px;"><span style="color:#006400">Antibiotic class codes on the y-axis are as follows:<br> | | <br> <span style="font-family:roboto;font-size:14px;"><span style="color:#006400">Antibiotic class codes on the y-axis are as follows:<br> |
| MC: macrolide, LM: lincosamide, TM: trimethoprim, TC: tetracycline,<br> | | MC: macrolide, LM: lincosamide, TM: trimethoprim, TC: tetracycline,<br> |
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| <li><span style="font-family:roboto;font-size:16px;">Patel, Robin. "Biofilms and antimicrobial resistance." Clinical Orthopaedics and Related Research<sup>®</sup> 437 (2005): 41-47.</span></li> | | <li><span style="font-family:roboto;font-size:16px;">Patel, Robin. "Biofilms and antimicrobial resistance." Clinical Orthopaedics and Related Research<sup>®</sup> 437 (2005): 41-47.</span></li> |
| <li><span style="font-family:roboto;font-size:16px;">Oechslin, Frank. "Resistance development to bacteriophages occurring during bacteriophage therapy." Viruses 10.7 (2018): 351.</span></li> | | <li><span style="font-family:roboto;font-size:16px;">Oechslin, Frank. "Resistance development to bacteriophages occurring during bacteriophage therapy." Viruses 10.7 (2018): 351.</span></li> |
− | <li><span style="font-family:roboto;font-size:16px;">Fig. 1: https://www.euro.who.int/__data/assets/pdf_file/0005/348224/Fact-sheet-SDG-AMR-FINAL-07-09-2017.pdf?ua=1</span></li> | + | <li><span style="font-family:roboto;font-size:16px;">https://www.euro.who.int/__data/assets/pdf_file/0005/348224/Fact-sheet-SDG-AMR-FINAL-07-09-2017.pdf?ua=1</span></li> |
− | <li><span style="font-family:roboto;font-size:16px;">Fig. 2: http://dbtindia.gov.in/sites/default/files/ScopingreportonAntimicrobialresistanceinIn dia.pdf</span></li> | + | <li><span style="font-family:roboto;font-size:16px;"> http://dbtindia.gov.in/sites/default/files/ScopingreportonAntimicrobialresistanceinIndia.pdf</span></li> |
− | <li><span style="font-family:roboto;font-size:16px;">Fig. 3: Science 29 Sep 2017:Vol. 357, Issue 6358, pp. 1350-1352, DOI:10.1126/science.aao1495</span></li> | + | <li><span style="font-family:roboto;font-size:16px;">Science 29 Sep 2017:Vol. 357, Issue 6358, pp. 1350-1352, DOI:10.1126/science.aao1495</span></li> |
| + | <li><span style="font-family:roboto;font-size:16px;">Van Epps, A., Blaney, L. Antibiotic Residues in Animal Waste: Occurrence and Degradation in Conventional Agricultural Waste Management Practices. Curr Pollution Rep 2, 135–155 (2016). https://doi.org/10.1007/s40726-016-0037-1</span></li> |
| <li><span style="font-family:roboto;font-size:16px;">Van Epps, A., Blaney, L. Antibiotic Residues in Animal Waste: Occurrence and Degradation in Conventional Agricultural Waste Management Practices. Curr Pollution Rep 2, 135–155 (2016). https://doi.org/10.1007/s40726-016-0037-1</span></li> | | <li><span style="font-family:roboto;font-size:16px;">Van Epps, A., Blaney, L. Antibiotic Residues in Animal Waste: Occurrence and Degradation in Conventional Agricultural Waste Management Practices. Curr Pollution Rep 2, 135–155 (2016). https://doi.org/10.1007/s40726-016-0037-1</span></li> |
− | <li><span style="font-family:roboto;font-size:16px;">Fig. 4: Van Epps, A., Blaney, L. Antibiotic Residues in Animal Waste: Occurrence and Degradation in Conventional Agricultural Waste Management Practices. Curr Pollution Rep 2, 135–155 (2016). https://doi.org/10.1007/s40726-016-0037-1</span></li>
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| <li><span style="font-family:roboto;font-size:16px;">Landers, T. F., Cohen, B., Wittum, T. E., & Larson, E. L. (2012). A review of antibiotic use in food animals: perspective, policy, and potential. Public health reports (Washington, D.C.: 1974), 127(1), 4–22. https://doi.org/10.1177/003335491212700103</span></li> | | <li><span style="font-family:roboto;font-size:16px;">Landers, T. F., Cohen, B., Wittum, T. E., & Larson, E. L. (2012). A review of antibiotic use in food animals: perspective, policy, and potential. Public health reports (Washington, D.C.: 1974), 127(1), 4–22. https://doi.org/10.1177/003335491212700103</span></li> |
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| </div><div class="clear"></div> | | </div><div class="clear"></div> |
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− | <p style="font-family:roboto;font-size:16px;"><br><span style="color:#f88379"><b>Fig 1 : </b>We would then be left with excreta free of both the antibiotics as well our engineered bacteria and could then be processed and used as safe manure. For proof of concept, we will initially target sulfonamides present in the poultry waste.</span><br><br> After the above process, cell lysate and enzymes could be found in the supernatant. Also to keep the temperature stable and suitable for enzyme action, the tank would be partially buried underground.<br><br> | + | <p style="font-family:roboto;font-size:16px;"><br><span style="color:#f88379"><b>Fig : </b>We would then be left with excreta free of both the antibiotics as well our engineered bacteria and could then be processed and used as safe manure. For proof of concept, we will initially target sulfonamides present in the poultry waste.</span><br><br> After the above process, cell lysate and enzymes could be found in the supernatant. Also to keep the temperature stable and suitable for enzyme action, the tank would be partially buried underground.<br><br> |
| <span style="font-family:roboto;font-size:18px;"><span style="color:#f88379"><b>How much do we degrade?</b></span></span><br><br> | | <span style="font-family:roboto;font-size:18px;"><span style="color:#f88379"><b>How much do we degrade?</b></span></span><br><br> |
| <span style="font-family:roboto;font-size:16px;">No biological reaction is 100% complete and thus we cannot really say that we can totally eliminate the antibiotics in the environment. But we can degrade antibiotics to such an extent that natural selection would start to favour the strains that are not resistant in the environment. Such limits for various antibiotics have been predicted and are termed as the proposed no effect concentrations (PNEC)<sup>1, 2</sup>. At these concentrations, the antibiotics in the environment would be so low that the gain of function mutation of AMR would be essentially rendered vestigial and would almost completely be eliminated by natural selection.</span><br><br> | | <span style="font-family:roboto;font-size:16px;">No biological reaction is 100% complete and thus we cannot really say that we can totally eliminate the antibiotics in the environment. But we can degrade antibiotics to such an extent that natural selection would start to favour the strains that are not resistant in the environment. Such limits for various antibiotics have been predicted and are termed as the proposed no effect concentrations (PNEC)<sup>1, 2</sup>. At these concentrations, the antibiotics in the environment would be so low that the gain of function mutation of AMR would be essentially rendered vestigial and would almost completely be eliminated by natural selection.</span><br><br> |
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| <br> | | <br> |
− | <p style="font-family:roboto;font-size:16px;"><span style="color:purple"><b>Fig 1:</b></span> This Module involves degradation of sulfonamides in the animal excreta. | + | <p style="font-family:roboto;font-size:16px;"><span style="color:purple"><b>Fig :</b></span> This Module involves degradation of sulfonamides in the animal excreta. |
| For this purpose the genes <i>sulX</i> and <i>sulR</i> | | For this purpose the genes <i>sulX</i> and <i>sulR</i> |
| were integrated downstream to a medium strength expression system. The genes <i>sulX</i> and <i>sulR</i> are a two component flavin dependent monooxygenase system present in <i>Microbacterium</i> sp. CJ77. | | were integrated downstream to a medium strength expression system. The genes <i>sulX</i> and <i>sulR</i> are a two component flavin dependent monooxygenase system present in <i>Microbacterium</i> sp. CJ77. |
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| <img src="https://static.igem.org/mediawiki/2020/e/e3/T--IISER-Tirupati_India--Poster_letsjustkeeponaddingstuffonthelastdaydefinitelyagoodidea.png " align="top"> | | <img src="https://static.igem.org/mediawiki/2020/e/e3/T--IISER-Tirupati_India--Poster_letsjustkeeponaddingstuffonthelastdaydefinitelyagoodidea.png " align="top"> |
| </div><br><div class="clear"></div> | | </div><br><div class="clear"></div> |
− | <p style="font-family:roboto;font-size:16px;"><span style="color:purple"><b>Fig 2:</b></span> Schematic depiction of two component flavin dependent monooxygenase system. | + | <p style="font-family:roboto;font-size:16px;"><span style="color:purple"><b>Fig :</b></span> Schematic depiction of two component flavin dependent monooxygenase system. |
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| </p><br><br> | | </p><br><br> |
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− | <p style="font-family:roboto;font-size:16px;"><span style="color:purple"><b>Fig 3:</b></span> Module-2 involves reducing conjugation thereby reducing horizontal gene transfer (HGT). To achieve this, the genes <i>traS</i> and <i>traT</i> are integrated downstream to a medium strength expression system in a F- chassis. The simultaneous expression of these genes in <i>E. coli</i> in a detergent free media has shown to reduce conjugation by 33,000 times. The genes <i>traS</i> and <i>traT</i> | + | <p style="font-family:roboto;font-size:16px;"><span style="color:purple"><b>Fig :</b></span> Module-2 involves reducing conjugation thereby reducing horizontal gene transfer (HGT). To achieve this, the genes <i>traS</i> and <i>traT</i> are integrated downstream to a medium strength expression system in a F- chassis. The simultaneous expression of these genes in <i>E. coli</i> in a detergent free media has shown to reduce conjugation by 33,000 times. The genes <i>traS</i> and <i>traT</i> |
| help in surface exclusion and are carried naturally by the <i>E. coli</i> F factor. | | help in surface exclusion and are carried naturally by the <i>E. coli</i> F factor. |
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− | <p style="font-family:roboto;font-size:16px;"><span style="color:purple"><b>Fig 4:</b></span> Module 3 involves designing an user modulated ‘kill switch’ to induce cell death in the engineered bacteria. For this, we take use of the arabinose inducible promoter. The reason for choice of this promoter could be attributed to the high expression, tight regulation in presence of glucose and a cost-effective inducer. The toxin used downstream to this promoter is the bovine pancreatic <i>DNaseI</i> which is a well characterized endonuclease. The circuit incorporates the <i>araC</i> gene which is expressed in the opposite direction to that of the <i>DNaseI.</i> Both Plasmids will contain the kill switch to ensure biosafety. | + | <p style="font-family:roboto;font-size:16px;"><span style="color:purple"><b>Fig :</b></span> Module 3 involves designing an user modulated ‘kill switch’ to induce cell death in the engineered bacteria. For this, we take use of the arabinose inducible promoter. The reason for choice of this promoter could be attributed to the high expression, tight regulation in presence of glucose and a cost-effective inducer. The toxin used downstream to this promoter is the bovine pancreatic <i>DNaseI</i> which is a well characterized endonuclease. The circuit incorporates the <i>araC</i> gene which is expressed in the opposite direction to that of the <i>DNaseI.</i> Both Plasmids will contain the kill switch to ensure biosafety. |
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| </p><br> | | </p><br> |
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| <span style="font-family:roboto; font-size:18px; color:#93225d"><b>Proposed Sulphonamide Degradation Mechanism:</b></span><br><br><div class="borderpurple"><img src="https://static.igem.org/mediawiki/2020/d/da/T--IISER-Tirupati_India--Poster_sdm.png" align="top"></div><br><br> | | <span style="font-family:roboto; font-size:18px; color:#93225d"><b>Proposed Sulphonamide Degradation Mechanism:</b></span><br><br><div class="borderpurple"><img src="https://static.igem.org/mediawiki/2020/d/da/T--IISER-Tirupati_India--Poster_sdm.png" align="top"></div><br><br> |
| <span style="font-family:roboto; font-size:16px;"><span style="color:#93225d"><b>Fig :</b></span>Proposed sulfonamide degradation mechanism for the initial cleavage reaction mediated by <i>sulX </i>(sulphonamide monooxygenase) and <i>sulR </i>(flavin reductase).<sup>1</sup></span><br><br> | | <span style="font-family:roboto; font-size:16px;"><span style="color:#93225d"><b>Fig :</b></span>Proposed sulfonamide degradation mechanism for the initial cleavage reaction mediated by <i>sulX </i>(sulphonamide monooxygenase) and <i>sulR </i>(flavin reductase).<sup>1</sup></span><br><br> |
− | <p style="font-family:roboto;font-size:16px;">Below set of equations can be used in any situation as it takes care of the fact that our enzyme is working in a non ideal medium.</p><br><span style="font-family:roboto; font-size:18px; color:#93225d"><b>Ordered bi-bi Mechanism:</b></span><br><div class="child2"><img src="https://static.igem.org/mediawiki/2020/c/c2/T--IISER-Tirupati_India--Poster_degradation10.png" align="top"></div><div class="clear"></div><br> | + | <p style="font-family:roboto;font-size:16px;">Below set of equations can be used in any situation as it takes care of the fact that our enzyme is working in a non ideal medium.</p><br><span style="font-family:roboto; font-size:18px; color:#93225d"><b>Ordered bi-bi Mechanism:</b></span><br><br><div class="borderpurple"><img src="https://static.igem.org/mediawiki/2020/3/3e/T--IISER-Tirupati_India--Poster_letsjustkeeponaddingstuffonthelastdaydefinitelyagoodideajustonemore.png" align="top"></div><div class="clear"></div><br><div class="child2"><img src="https://static.igem.org/mediawiki/2020/c/c2/T--IISER-Tirupati_India--Poster_degradation10.png" align="top"></div><div class="clear"></div><br> |
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− | <span style="font-family:roboto; font-size:18px; color:#93225d"><b>Random Bisubstrate Mechanism:</b></span><br> | + | <span style="font-family:roboto; font-size:18px; color:#93225d"><b>Random Bisubstrate Mechanism:</b></span><br><br><div class="borderpurple"><img src="https://static.igem.org/mediawiki/2020/5/54/T--IISER-Tirupati_India--Poster_letsjustkeeponaddingstuffonthelastdaydefinitelyagoodideaanotherseriously.png" align="top"></div><div class="clear"></div><br> |
| <div class="child2"><img src="https://static.igem.org/mediawiki/2020/3/34/T--IISER-Tirupati_India--Poster_degradation11.png" align="top"></div><div class="clear"></div> | | <div class="child2"><img src="https://static.igem.org/mediawiki/2020/3/34/T--IISER-Tirupati_India--Poster_degradation11.png" align="top"></div><div class="clear"></div> |
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| <p style="font-family:roboto;font-size:18px;"><span style="color:#93225d"><b>Ordered bi-bi Mechanism:</b></span></p><br> | | <p style="font-family:roboto;font-size:18px;"><span style="color:#93225d"><b>Ordered bi-bi Mechanism:</b></span></p><br> |
| <div class="borderpurple"><img src="https://static.igem.org/mediawiki/2020/1/14/T--IISER-Tirupati_India--Poster_degradation2.png" align="top"></div><div class="clear"></div><br> | | <div class="borderpurple"><img src="https://static.igem.org/mediawiki/2020/1/14/T--IISER-Tirupati_India--Poster_degradation2.png" align="top"></div><div class="clear"></div><br> |
− | <div class="borderpurple"><img src="https://static.igem.org/mediawiki/2020/3/3e/T--IISER-Tirupati_India--Poster_letsjustkeeponaddingstuffonthelastdaydefinitelyagoodideajustonemore.png" align="top"></div><div class="clear"></div><br>
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| <div class="borderpurple"><img src="https://static.igem.org/mediawiki/2020/f/f5/T--IISER-Tirupati_India--Poster_degradation3.png" align="top"></div> | | <div class="borderpurple"><img src="https://static.igem.org/mediawiki/2020/f/f5/T--IISER-Tirupati_India--Poster_degradation3.png" align="top"></div> |
| <div class="clear"></div><br> | | <div class="clear"></div><br> |
− | <div class="borderpurple"><img src="https://static.igem.org/mediawiki/2020/5/54/T--IISER-Tirupati_India--Poster_letsjustkeeponaddingstuffonthelastdaydefinitelyagoodideaanotherseriously.png" align="top"></div><div class="clear"></div><br>
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| <p style="font-family:roboto;font-size:18px;"><span style="color:#93225d"><b>Key Inferences:</b></span></p><ul> | | <p style="font-family:roboto;font-size:18px;"><span style="color:#93225d"><b>Key Inferences:</b></span></p><ul> |
| <li><span style="font-family:roboto;font-size:16px;">Decreasing the <span style="color:#93225d">𝛂<sub>2</sub></span> value increases the time taken to degrade antibiotics.</span></li> | | <li><span style="font-family:roboto;font-size:16px;">Decreasing the <span style="color:#93225d">𝛂<sub>2</sub></span> value increases the time taken to degrade antibiotics.</span></li> |
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| <div class="title"> <span style="color:#93225d">Horizontal Gene Transfer Reduction Model</span> </div> | | <div class="title"> <span style="color:#93225d">Horizontal Gene Transfer Reduction Model</span> </div> |
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− | <!--Section text---><div class="text"><span style="font-family:roboto;font-size:18px;"><span style="color:#93225d"><b>Transformation:</b></span></span><br><div class="borderchild4"><img src="https://static.igem.org/mediawiki/2020/8/8f/T--IISER-Tirupati_India--Poster_hgt1.png" align="top"></div> | + | <!--Section text---><div class="text"><span style="font-family:roboto;font-size:18px;"><span style="color:#93225d"><b>Transformation:</b></span></span><br><br> |
| + | <div class="borderchild4"><img src="https://static.igem.org/mediawiki/2020/7/77/T--IISER-Tirupati_India--Poster_longestposterever.png" align="top"></div> |
| + | <div class="clear"></div><br><div class="borderchild4"><img src="https://static.igem.org/mediawiki/2020/8/86/T--IISER-Tirupati_India--Poster_MIjustwon.png" align="top"></div> |
| + | <div class="clear"></div><br><div class="borderchild4"><img src="https://static.igem.org/mediawiki/2020/8/8f/T--IISER-Tirupati_India--Poster_hgt1.png" align="top"></div> |
| <div class="clear"></div><br><span style="font-family:roboto;font-size:18px;"><span style="color:#93225d"><b>Key inferences:</b></span></span><ul> | | <div class="clear"></div><br><span style="font-family:roboto;font-size:18px;"><span style="color:#93225d"><b>Key inferences:</b></span></span><ul> |
| <li><span style="font-family:roboto;font-size:16px;">The double gene system buys us more time for degrading the antibiotics.</span></li> | | <li><span style="font-family:roboto;font-size:16px;">The double gene system buys us more time for degrading the antibiotics.</span></li> |
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| <div class="clear"></div><br> | | <div class="clear"></div><br> |
| | | |
− | <span style="font-family:roboto;font-size:18px;"><span style="color:#93225d"><b>Conjugation:</b></span></span><br><br><div class="borderchild4"><img src="https://static.igem.org/mediawiki/2020/8/8a/T--IISER-Tirupati_India--Poster_hgt2.png" align="top"></div> | + | <span style="font-family:roboto;font-size:18px;"><span style="color:#93225d"><b>Conjugation:</b></span></span><br><br><div class="borderchild4"><img src="https://static.igem.org/mediawiki/2020/8/87/T--IISER-Tirupati_India--Poster_nowgettingscoldedathome.png" align="top"></div> |
| + | <div class="clear"></div><br><div class="borderchild4"><img src="https://static.igem.org/mediawiki/2020/8/8a/T--IISER-Tirupati_India--Poster_hgt2.png" align="top"></div> |
| <div class="clear"></div> | | <div class="clear"></div> |
| <br><span style="font-family:roboto;font-size:18px;"><span style="color:#93225d"><b>Key Inferences:</b></span><ul> | | <br><span style="font-family:roboto;font-size:18px;"><span style="color:#93225d"><b>Key Inferences:</b></span><ul> |
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| <li><span style="font-family:roboto;font-size:16px;">The two proteins TraS and TraT reduce the formation of transconjugants to such an extent that with these initial conditions, it takes about 50 hrs to form one mutant in a 100L slurry.</span></li> | | <li><span style="font-family:roboto;font-size:16px;">The two proteins TraS and TraT reduce the formation of transconjugants to such an extent that with these initial conditions, it takes about 50 hrs to form one mutant in a 100L slurry.</span></li> |
| <li><span style="font-family:roboto;font-size:16px;">The concentration of transconjugants rises extremely slowly because of low growth rate, low mating pair formation rate and high mating pair breaking rate.</span></li></ul> | | <li><span style="font-family:roboto;font-size:16px;">The concentration of transconjugants rises extremely slowly because of low growth rate, low mating pair formation rate and high mating pair breaking rate.</span></li></ul> |
| + | <br> |
| + | <span style="font-family:roboto;font-size:18px;"><span style="color:#93225d"><b>Transduction:</b></span></span><br><br><div class="borderchild4"><img src="https://static.igem.org/mediawiki/2020/f/f8/T--IISER-Tirupati_India--Poster_justafewmore.png" align="top"></div> |
| + | <div class="clear"></div><br><div class="borderchild4"><img src="https://static.igem.org/mediawiki/2020/b/b7/T--IISER-Tirupati_India--Poster_justafewmoremaybe.png" align="top"></div> |
| + | <div class="clear"></div><br> |
| + | <span style="font-family:roboto;font-size:16px;">The concentration of AMR mutants would be calculated by the following equations:<br> |
| + | For single gene system: |
| + | </span> |
| <br><br> | | <br><br> |
− | | + | <div class="subsection full"><img src="https://static.igem.org/mediawiki/2020/9/9a/T--IISER-Tirupati_India--Poster_lastbutone.png" align="top"></div><div class="clear"></div><br> |
− | <span style="font-family:roboto;font-size:18px;"><span style="color:#93225d"><b>Transduction:</b></span></span><br><div class="borderchild4"><img src="https://static.igem.org/mediawiki/2020/7/71/T--IISER-Tirupati_India--Poster_hgt3.png" align="top"></div><div class="clear"></div> | + | <span style="font-family:roboto;font-size:16px;">For double gene system, the term b_fr_AMR_en is squared as there are two plasmids that the virus has to pick up.<br> |
| + | Therefore, the double gene system:</span><br><br> |
| + | <div class="subsection full"><img src="https://static.igem.org/mediawiki/2020/c/c8/T--IISER-Tirupati_India--Poster_last.png" align="top"></div><div class="clear"></div><br> |
| + | <div class="borderchild4"><img src="https://static.igem.org/mediawiki/2020/7/71/T--IISER-Tirupati_India--Poster_hgt3.png" align="top"></div><div class="clear"></div><br> |
| <span style="font-family:roboto;font-size:18px;"><span style="color:#93225d"><b>Key inferences:</b></span></span><ul> | | <span style="font-family:roboto;font-size:18px;"><span style="color:#93225d"><b>Key inferences:</b></span></span><ul> |
| <li><span style="font-family:roboto;font-size:16px;">The AMR mutant does not persist in the environment, but the cells harboring the virus particles with the AMR genes are stabilizing in the environment and so are the virus particles.</span></li></ul> | | <li><span style="font-family:roboto;font-size:16px;">The AMR mutant does not persist in the environment, but the cells harboring the virus particles with the AMR genes are stabilizing in the environment and so are the virus particles.</span></li></ul> |
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| <!--Section text---> | | <!--Section text---> |
| <div class="text"> | | <div class="text"> |
| + | <span style="font-family:roboto;font-size:18px;"><span style="color:#93225d"><b>Working:</b></span></span><br><br> |
| + | <div class="borderchild4"><img src="https://static.igem.org/mediawiki/2020/b/bb/T--IISER-Tirupati_India--Poster_okayiliedtherearemore.png" align="top"></div><br><br> |
| + | |
| + | <span style="font-family:roboto;font-size:18px;"><span style="color:#93225d"><b>Production of DNAse I protein:</b></span></span><br><br> |
| + | <div class="child2"><img src="https://static.igem.org/mediawiki/2020/3/33/T--IISER-Tirupati_India--Poster_okayiliedtherearemorepart4.png" align="top"></div><br><br> |
| + | |
| + | <span style="font-family:roboto;font-size:18px;"><span style="color:#93225d"><b>DNA Degradation:</b></span></span><br><br> |
| + | <div class="child2"><img src="https://static.igem.org/mediawiki/2020/b/bc/T--IISER-Tirupati_India--Poster_okayiliedtherearemorepart3.png" align="top"></div><br><br> |
| | | |
| <span style="font-family:roboto;font-size:18px;"><span style="color:#93225d"><b>Plot Ara C vs. Time:</b></span></span><br><span style="font-family:roboto;font-size:16px;">To find excess arabinose molecules in system = | | <span style="font-family:roboto;font-size:18px;"><span style="color:#93225d"><b>Plot Ara C vs. Time:</b></span></span><br><span style="font-family:roboto;font-size:16px;">To find excess arabinose molecules in system = |
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| | | |
| <div class="borderchild4"><img src="https://static.igem.org/mediawiki/2020/8/81/T--IISER-Tirupati_India--Poster_time3d.png" align="top"></div> | | <div class="borderchild4"><img src="https://static.igem.org/mediawiki/2020/8/81/T--IISER-Tirupati_India--Poster_time3d.png" align="top"></div> |
| + | <div class="clear"></div><br><div class="borderchild4"><img src="https://static.igem.org/mediawiki/2020/6/65/T--IISER-Tirupati_India--Poster_okayiliedtherearemorepart2.png" align="top"></div> |
| <div class="clear"></div><br><span style="font-family:roboto;font-size:16px;"><span style="color:#93225d"><b>Inference:</b></span> The maximum number of c-AMP molecules that can be present in a cell is 89727.8 molecules per cell.</span><br><br> | | <div class="clear"></div><br><span style="font-family:roboto;font-size:16px;"><span style="color:#93225d"><b>Inference:</b></span> The maximum number of c-AMP molecules that can be present in a cell is 89727.8 molecules per cell.</span><br><br> |
| | | |
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| <div class="borderpink"><img src="https://static.igem.org/mediawiki/2020/e/e5/T--IISER-Tirupati_India--Poster_manavarachana.png" align="top"></div><br><br> | | <div class="borderpink"><img src="https://static.igem.org/mediawiki/2020/e/e5/T--IISER-Tirupati_India--Poster_manavarachana.png" align="top"></div><br><br> |
| | | |
− | | + | <span style="font-family:roboto;font-size:18px;"><span style="color:#FF69B4"><b>Interaction with Undergraduate Students at |
| + | Yeshwant Mahavidyalaya Nanded</b></span></span><br><br> |
| + | <div class="borderpink"><img src="https://static.igem.org/mediawiki/2020/4/40/T--IISER-Tirupati_India--Poster_motivationdomujhe.png" align="top"></div><br><br> |
| | | |
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| <li><span style=font-family: roboto; font-size:16px;color:#003366;”><span style="color:#003366"> Even though our project faced several difficulties this year, we were successfully able to complete the first phase of The Engineering Design Cycle involving Research, Imagine and Design with certain preliminary Build. Additionally, in a <b>year-wide collaboration with iGEM IONIS Paris</b> we could successfully develop clones with our 'kill switch' and are yet to characterize them. | | <li><span style=font-family: roboto; font-size:16px;color:#003366;”><span style="color:#003366"> Even though our project faced several difficulties this year, we were successfully able to complete the first phase of The Engineering Design Cycle involving Research, Imagine and Design with certain preliminary Build. Additionally, in a <b>year-wide collaboration with iGEM IONIS Paris</b> we could successfully develop clones with our 'kill switch' and are yet to characterize them. |
| </span></li> | | </span></li> |
| + | <li><span style=font-family: roboto; font-size:16px;color:#003366;”><span style="color:#003366"><b>Won Gold</b> in iGEM World Wide Virtual meet up hosted by Parisian Teams for Best Pitch Presentation.</span></li> |
| <li><span style=font-family: roboto; font-size:16px;color:#003366;”><span style="color:#003366"> <b>As a team, we wrote three reviews one of which summarized our project in detail.</b> Moreover, not just writing, we also got to peer review some of the other articles for the journal and appreciate the work of other teams in the competition. | | <li><span style=font-family: roboto; font-size:16px;color:#003366;”><span style="color:#003366"> <b>As a team, we wrote three reviews one of which summarized our project in detail.</b> Moreover, not just writing, we also got to peer review some of the other articles for the journal and appreciate the work of other teams in the competition. |
| </span></li> | | </span></li> |
| <li><span style=font-family: roboto; font-size:16px;color:#003366;”><span style="color:#003366"> <b>Delivered a promising technological design and implementation</b> that will be faster and efficient than all the technology currently used to tackle Antibiotic Pollution as bacteria grow and metabolize very quickly. | | <li><span style=font-family: roboto; font-size:16px;color:#003366;”><span style="color:#003366"> <b>Delivered a promising technological design and implementation</b> that will be faster and efficient than all the technology currently used to tackle Antibiotic Pollution as bacteria grow and metabolize very quickly. |
− | </span></li></ul> | + | </span></li> |
| + | |
| + | </ul> |
| | | |
| | | |