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| | <h6><b>Improve</b></h6> | | <h6><b>Improve</b></h6> |
| − | <p><span style="font-weight: 400;"><strong>Improvement upon part <a href="http://parts.igem.org/Part:BBa_K2299000">BBa_K2299000</a></strong></span></p>
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| − | <p><span style="font-weight: 400;">The 2018 iGEM NYU Abu Dhabi designed a BioBrick that includes a gene that can be targeted for species-specific detection of </span><em><span style="font-weight: 400;">Vibrio cholera.</span></em></p>
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| − | <p><span style="font-weight: 400;">Part </span><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2808000"><span style="font-weight: 400;">BBa_K2808000</span></a><span style="font-weight: 400;"> was designed by including N-acetyl glucosamine-binding protein A (gbpA) gene for the detection of</span><em><span style="font-weight: 400;"> Vibrio cholera. </span></em><span style="font-weight: 400;">The gbpA gene in Vibrio cholerae codes for the N-acetyl glucosamine-binding protein A (GbpA), which is a chitin-binding protein involved in the attachment of </span><em><span style="font-weight: 400;">V. cholerae</span></em><span style="font-weight: 400;"> to environmental chitin surfaces and human intestinal cells. The gbpA gene has been found to be consistently present and highly conserved in </span><em><span style="font-weight: 400;">V. cholerae</span></em><span style="font-weight: 400;">. GbpA has been used as a target gene for the species-specific detection of </span><em><span style="font-weight: 400;">V. cholerae</span></em><span style="font-weight: 400;"> O1, O139, Non-O1/Non-O139.</span></p>
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| − | <h4><span style="font-weight: 400;">Instead of the 39 bp sequence found in CTX RstA gene that is used by </span><a href="http://parts.igem.org/Part:BBa_K2299000"><span style="font-weight: 400;">BBa_K2299000</span></a><span style="font-weight: 400;">, part </span><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2808000"><span style="font-weight: 400;">BBa_K2808000</span></a><span style="font-weight: 400;"> uses gbpA gene for the detection of </span><em><span style="font-weight: 400;">V. cholera. </span></em><span style="font-weight: 400;">The advantages of using gbpA as a gene target compared to ctxA are various such as ctxA is involved in the production of the cholera toxin production which requires special safety precautions during bacterial transformation, lab experiments, shipping reagents, customs, etc. whereas the use of gbpA solves these problems as gbpA is not directly involved in the production of cholera toxin.</span></h4>
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| − | <h4><u>gbpA BioBrick double digestion reaction</u></h4>
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| − | <p><img class="center2" src="https://static.igem.org/mediawiki/2018/b/bf/T--NYU_Abu_Dhabi--Parts3.png" height="400" /></p>
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| − | <p>Figure 1: Agarose gel electrophoresis showing double digestion of the gbpA BioBrick <a href="http://parts.igem.org/Part:BBa_K2808000" target="_blank" rel="noopener">BBA_K2808000</a> (gbpA gene (1458 bp) inserted into the pSB1C3 backbone (2070 bp)) with EcoRI and PstI. (Lane 1) 500 bp DNA ladder (Bio-Rad); (Lane 2) gbpA BioBrick; (Lane 3) gbpA BioBrick digested with EcoRI and PstI.</p>
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| − | <p> <br /><u>BioBricks PCR amplification</u></p>
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| − | <p><img class="center" src="https://static.igem.org/mediawiki/2018/9/9d/T--NYU_Abu_Dhabi--Parts1.png" width="300" /> </p>
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| − | <p>Figure 1: Agarose gel electrophoresis showing PCR amplification of: gbpA BioBrick <a href="http://parts.igem.org/Part:BBa_K2808000" target="_blank" rel="noopener">BBA_K2808000</a>; hipO <a href="http://parts.igem.org/Part:BBa_K2808001" target="_blank" rel="noopener">BBA_K2808001</a>; invA <a href="http://parts.igem.org/Part:BBa_K2808002" target="_blank" rel="noopener">BBA_K2808002</a>; lmo0733 <a href="http://parts.igem.org/Part:BBa_K2808003" target="_blank" rel="noopener">BBA_K2808003</a>. (Lane 1) 500 bp DNA ladder (Bio-Rad); (Lane 2) gbpA BioBrick; (Lane 3) negative control for gbpA BioBrick; (Lane 4) hipO BioBrick; (Lane 5) negative control for hipO BioBrick; (Lane 6) invA BioBrick; (Lane 7) negative control for invA BioBrick; (Lane 8) lmo0733 BioBrick; (Lane 9) negative control for lmo0733 BioBrick. Lanes 2, 4, 6, and 8 show bands around 1019 bp, 203 bp, 818 bp, and 403 bp respectively as expected for the PCR products of the gbpA, hipO, invA, and lmo0733 BioBricks respectively.</p>
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| − | <p> </p>
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| − | <h2> </h2>
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| − | <p> </p>
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| − | <p><br /><br /><br /></p>
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