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| − | <link rel="stylesheet" href="https://www.ckmoban.com/igem/1382/css/mmenu.css">
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| − | <link rel="stylesheet" href="https://www.ckmoban.com/igem/1382/css/responsive.css">
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| − | <link rel="stylesheet" href="https://www.ckmoban.com/igem/1382/layerslider/css/layerslider.css">
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| − | <link rel="stylesheet" href="https://www.ckmoban.com/igem/1382/css/twentytwenty.css">
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| − | <link rel="stylesheet" href="https://www.ckmoban.com/igem/1382/css/test.css">
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| − | <link rel="shortcut icon" href="img/favicon.png">
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| − | <h1>Description</h1> | + | <h1>Design</h1> |
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| − | <p><span style=";font-family:'Times New Roman';font-size:16px">Our project has two parts: biosensor and biosorption</span></p><h1><strong><span style="font-family: 'Times New Roman';font-size: 16px">1. The biosensor</span></strong></h1><p><span style=";font-family:'Times New Roman';font-size:16px">In the design of our biosensor, we used the part </span><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">BBa_J33201</span></span><span style=";font-family:'Times New Roman';font-size:16px"> registered By Edinburgh University in 2006 as the basis to build our biosensor, and installed a strong RBS and GFP reporter gene protein downstream of the part to build our biosensor.</span></p><p><span style=";font-family:'Times New Roman';font-size:16px"> </span></p><p><span style=";font-family:'Times New Roman';font-size:16px">The reason for this construction is that we got inspiration from the part </span><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">BBa�_J33203</span></span><span style=";font-family:'Times New Roman';font-size:16px"> registered by the Edinburg Team in 2006, so we did not add terminator in the gene pathway when building this Biosensor.</span></p><p><span style=";font-family:'Times New Roman';font-size:16px"> </span></p><p><span style=";font-family:'Times New Roman';font-size:16px">Fortunately, we found that our biosensor could work normally and had a sensitive detection threshold for arsenic ions in water.</span></p><p><span style=";font-family:'Times New Roman';font-size:16px"> </span></p><p><span style=";font-family:'Times New Roman';font-size:16px">At the same time, we also characterized </span><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">J33201</span></span><span style=";font-family:'Times New Roman';font-size:16px">, an excellent part, and added more detailed and valuable data for the part.</span></p><p><span style=";font-family:'Times New Roman';font-size:16px">Our genetic pathway is designed as follows</span> </p><p><img alt="Design(图1)" width="419" height="207" src="/public/plugins/Ueditor/themes/default/images/spacer.gif" word_img="file:///C:\Users\ADMINI~1\AppData\Local\Temp\ksohtml\wps1E7D.tmp.jpg" style="max-width:100%!important;height:auto;background:url(/public/plugins/Ueditor/lang/zh-cn/images/localimage.png) no-repeat center center;border:1px solid #ddd"><span style=";font-family:'Times New Roman';font-size:16px"> </span></p><p><a href="http://parts.igem.org/wiki/images/e/e4/T--Worldsher-wuhan-design_1.png"><span style="text-decoration:underline;"><span style="font-family: 楷体;color: rgb(0, 0, 255);font-size: 16px">http://parts.igem.org/wiki/images/e/e4/T--Worldsher-wuhan-design_1.png</span></span></a></p><p><span style=";font-family:'Times New Roman';font-size:16px"> </span></p><p><span style=";font-family:'Times New Roman';font-size:16px">J33201 is a part registered with the Edinburg Team in 2006, which includes an arsenic-sensitive promoter and </span><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">ArsR,</span></span><span style=";font-family:'Times New Roman';font-size:16px"> an auto-regulatory repressor from </span><em><span style="font-family: 'Times New Roman';font-size: 16px">E. coli.</span></em><span style=";font-family:'Times New Roman';font-size:16px"> On this basis, they constructed one of the biosensors, J3320 with a reporter gene of </span><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">lacZ</span></span><span style=";font-family:'Times New Roman';font-size:16px">, expressing galactosidase. </span></p><p><span style=";font-family:'Times New Roman';font-size:16px"> </span></p><p><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">LacZ</span></span><span style=";font-family:'Times New Roman';font-size:16px"> can react with substrate </span><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">x-gal</span></span><span style=";font-family:'Times New Roman';font-size:16px"> for colour development, yet we found in the preliminary experiment that this component was less sensitive to arsenic than expected, and the reaction time was very long.</span></p><p><span style=";font-family:'Times New Roman';font-size:16px"> </span></p><p><span style=";font-family:'Times New Roman';font-size:16px">So, we construct a biosensor expressing </span><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">GFP</span></span><span style=";font-family:'Times New Roman';font-size:16px">, and on this basis we characterize</span><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px"> J33201</span></span><span style=";font-family:'Times New Roman';font-size:16px"> again.</span></p><h1><strong><span style="font-family: 'Times New Roman';font-size: 16px">2.Biosorption</span></strong></h1><p><span style=";font-family:'Times New Roman';font-size:16px">Our biosorption consists of two components:</span></p><p><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">Metallothionein (MT)</span></span><span style=";font-family:'Times New Roman';font-size:16px"> plays an important role in bioremediation of heavy metals. </span><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">MT</span></span><span style=";font-family:'Times New Roman';font-size:16px"> is a kind of small molecular peptide rich in </span><em><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">cysteine (Cys-)</span></span></em><span style=";font-family:'Times New Roman';font-size:16px"> residues with low molecular weight. Its </span><em><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">Cys-</span></span></em><span style=";font-family:'Times New Roman';font-size:16px"> residues can adsorb many heavy metals such as lead, mercury and cadmium. However, since arsenic is a metalloid, these </span><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">MTs</span></span><span style=";font-family:'Times New Roman';font-size:16px"> have no specific adsorption on it. </span></p><p><span style=";font-family:'Times New Roman';font-size:16px"> </span></p><p><span style=";font-family:'Times New Roman';font-size:16px">Recently, scientists isolated and identified an </span><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">MT </span></span><span style=";font-family:'Times New Roman';font-size:16px">from a species of arsenic-tolerant Marine algae (</span><em><span style="font-family: 'Times New Roman';font-size: 16px">Fusus vesiculosus)</span></em><span style=";font-family:'Times New Roman';font-size:16px"> that, in vitro, demonstrated a high affinity for arsenic binding capacity.</span></p><p><span style=";font-family:'Times New Roman';font-size:16px"> </span></p><p><span style=";font-family:'Times New Roman';font-size:16px">In this project, we used the bacterial surface display technology to display </span><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">fMT</span></span><span style=";font-family:'Times New Roman';font-size:16px"> on the bacterial surface, and thus enhance the ability of absorbing arsenic.</span></p><p><span style=";font-family:'Times New Roman';font-size:16px"> </span></p><p><span style=";font-family:'Times New Roman';font-size:16px">Inspired by HUST-China team in 2014, the </span><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">fusion protein oprf-fMT</span></span><span style=";font-family:'Times New Roman';font-size:16px"> was constructed and heterogeneiously expressed by the </span><em><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">E. coli</span></span></em><span style=";font-family:'Times New Roman';font-size:16px"> expression system pET series plasmid.</span></p><p><span style=";font-family:'Times New Roman';font-size:16px"> </span></p><p><span style=";font-family:'Times New Roman';font-size:16px">Our design is as follows</span></p><p><img alt="Design(图2)" width="273" height="163" src="/public/plugins/Ueditor/themes/default/images/spacer.gif" word_img="file:///C:\Users\ADMINI~1\AppData\Local\Temp\ksohtml\wps1EBC.tmp.jpg" style="max-width:100%!important;height:auto;background:url(/public/plugins/Ueditor/lang/zh-cn/images/localimage.png) no-repeat center center;border:1px solid #ddd"><span style=";font-family:'Times New Roman';font-size:16px"> </span></p><p><a href="http://parts.igem.org/wiki/images/7/72/T--Worldsher-wuhan-design_2.png"><span style="text-decoration:underline;"><span style="font-family: 楷体;color: rgb(0, 0, 255);font-size: 16px">http://parts.igem.org/wiki/images/7/72/T--Worldsher-wuhan-design_2.png</span></span></a></p><p><span style=";font-family:'Times New Roman';font-size:16px"> </span></p><p><img alt="Design(图3)" width="336" height="233" src="/public/plugins/Ueditor/themes/default/images/spacer.gif" word_img="file:///C:\Users\ADMINI~1\AppData\Local\Temp\ksohtml\wps1ECD.tmp.jpg" style="max-width:100%!important;height:auto;background:url(/public/plugins/Ueditor/lang/zh-cn/images/localimage.png) no-repeat center center;border:1px solid #ddd"><span style=";font-family:'Times New Roman';font-size:16px"> </span></p><p><a href="http://parts.igem.org/wiki/images/3/3b/T--Worldsher-wuhan-design_3.png"><span style="text-decoration:underline;"><span style="font-family: 楷体;color: rgb(0, 0, 255);font-size: 16px">http://parts.igem.org/wiki/images/3/3b/T--Worldsher-wuhan-design_3.png</span></span></a></p><p><img alt="Design(图4)" width="449" height="198" src="/public/plugins/Ueditor/themes/default/images/spacer.gif" word_img="file:///C:\Users\ADMINI~1\AppData\Local\Temp\ksohtml\wps1ECE.tmp.jpg" style="max-width:100%!important;height:auto;background:url(/public/plugins/Ueditor/lang/zh-cn/images/localimage.png) no-repeat center center;border:1px solid #ddd"><span style=";font-family:'Times New Roman';font-size:16px"> </span></p><p><a href="http://parts.igem.org/wiki/images/9/9b/T-Worldsher-wuhan-design_4.png"><span style="text-decoration:underline;"><span style="font-family: 楷体;color: rgb(0, 0, 255);font-size: 16px">http://parts.igem.org/wiki/images/9/9b/T-Worldsher-wuhan-design_4.png</span></span></a></p><p><span style=";font-family:'Times New Roman';font-size:16px"> </span></p><p><span style=";font-family:'Times New Roman';font-size:16px">In addition, an important protein called </span><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">phytochelates (PC)</span></span><span style=";font-family:'Times New Roman';font-size:16px"> with a high affinity for arsenic has been found. PC is a natural peptide in plants and fungi, the general formula of the structure is</span><em><span style="font-family: 'Times New Roman';font-size: 16px"> </span></em><em><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">(</span></span></em><em><span style="text-decoration:underline;"><span style="font-family: 楷体;font-size: 16px">γ</span></span></em><em><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">-Glu-Cys-)n-Gly(n=1-11) </span></span></em><span style=";font-family:'Times New Roman';font-size:16px">. It was found that </span><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">PC</span></span><span style=";font-family:'Times New Roman';font-size:16px"> had a higher adsorption capacity for arsenic than </span><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">MT</span></span><span style=";font-family:'Times New Roman';font-size:16px">. </span><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">PC</span></span><span style=";font-family:'Times New Roman';font-size:16px"> is synthesized by </span><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">PC synthase (PCS)</span></span><span style=";font-family:'Times New Roman';font-size:16px"> with </span><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">glutathione (GSH)</span></span><span style=";font-family:'Times New Roman';font-size:16px"> as its substrate. </span></p><p><span style=";font-family:'Times New Roman';font-size:16px"> </span></p><p><span style=";font-family:'Times New Roman';font-size:16px">Previous studies have found that </span><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">PC synthase</span></span><span style=";font-family:'Times New Roman';font-size:16px"> from </span><em><span style="font-family: 'Times New Roman';font-size: 16px">Shizosomyces pombe</span></em><span style=";font-family:'Times New Roman';font-size:16px"> can be expressed in </span><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">SpPCS</span></span><span style=";font-family:'Times New Roman';font-size:16px"> in </span><em><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">E. coli</span></span></em><span style=";font-family:'Times New Roman';font-size:16px"> and thus enhance the adsorption ability of </span><em><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">E. coli</span></span></em><span style=";font-family:'Times New Roman';font-size:16px"> to arsenic.</span></p><p><span style=";font-family:'Times New Roman';font-size:16px"> </span></p><p><span style=";font-family:'Times New Roman';font-size:16px">Therefore, we artificially synthesized </span><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">ScPCS</span></span><span style=";font-family:'Times New Roman';font-size:16px"> genes (partial base changes to make them conform to RFC10 standard) and expressed them in pET expression system</span></p><p><span style=";font-family:'Times New Roman';font-size:16px"> </span></p><p><span style=";font-family:'Times New Roman';font-size:16px">Design is as follows</span></p><p><img alt="Design(图5)" width="262" height="82" src="/public/plugins/Ueditor/themes/default/images/spacer.gif" word_img="file:///C:\Users\ADMINI~1\AppData\Local\Temp\ksohtml\wps1EDE.tmp.jpg" style="max-width:100%!important;height:auto;background:url(/public/plugins/Ueditor/lang/zh-cn/images/localimage.png) no-repeat center center;border:1px solid #ddd"><span style=";font-family:楷体;font-size:16px"> </span></p><p><a href="http://parts.igem.org/wiki/images/4/40/T-Worldsher-wuhan-design_5.png"><span style="text-decoration:underline;"><span style="font-family: 楷体;color: rgb(0, 0, 255);font-size: 16px">http://parts.igem.org/wiki/images/4/40/T-Worldsher-wuhan-design_5.png</span></span></a></p><p><span style="text-decoration:underline;"><span style="font-family: 楷体;color: rgb(0, 0, 255);font-size: 16px"> </span></span></p><p><img alt="Design(图6)" width="273" height="223" src="/public/plugins/Ueditor/themes/default/images/spacer.gif" word_img="file:///C:\Users\ADMINI~1\AppData\Local\Temp\ksohtml\wps1EEF.tmp.jpg" style="max-width:100%!important;height:auto;background:url(/public/plugins/Ueditor/lang/zh-cn/images/localimage.png) no-repeat center center;border:1px solid #ddd"><span style=";font-family:楷体;font-size:16px"> </span></p><p><a href="http://parts.igem.org/wiki/images/9/9f/T-Worldsher-wuhan-design_6.png"><span style="text-decoration:underline;"><span style="font-family: 楷体;color: rgb(0, 0, 255);font-size: 16px">http://parts.igem.org/wiki/images/9/9f/T-Worldsher-wuhan-design_6.png</span></span></a></p><p><span style=";font-family:楷体;font-size:16px"> </span></p><p><img alt="Design(图7)" width="549" height="239" src="/public/plugins/Ueditor/themes/default/images/spacer.gif" word_img="file:///C:\Users\ADMINI~1\AppData\Local\Temp\ksohtml\wps1EF0.tmp.jpg" style="max-width:100%!important;height:auto;background:url(/public/plugins/Ueditor/lang/zh-cn/images/localimage.png) no-repeat center center;border:1px solid #ddd"><span style=";font-family:楷体;font-size:16px"> </span></p><p><a href="http://parts.igem.org/wiki/images/5/55/T-Worldsher-wuhan-design_7.png"><span style="text-decoration:underline;"><span style="font-family: 楷体;color: rgb(0, 0, 255);font-size: 16px">http://parts.igem.org/wiki/images/5/55/T-Worldsher-wuhan-design_7.png</span></span></a></p><p><span style=";font-family:'Times New Roman';font-size:16px"> </span></p><p><span style=";font-family:'Times New Roman';font-size:16px">We hope that the synergistic effect can be achieved by constructing the recombinant bacteria with adsorption within the bacteria and the adsorption using surface display, and thus we can remove arsenic ions from water more effectively.</span></p><p><span style=";font-family:'Times New Roman';font-size:16px"> </span></p><p><span style=";font-family:'Times New Roman';font-size:16px">In conclusion, we intend to build a biosensor to monitor the arsenic concentration in water and use the bioremediation device we built to remove arsenic ions in water.</span></p><p><br></p>
| + | <p><span style=";font-family:'Times New Roman';font-size:16px">Our project has two parts: biosensor and biosorption</span></p><h1><strong><span style="font-family: 'Times New Roman';font-size: 16px">1. The biosensor</span></strong></h1><p><span style=";font-family:'Times New Roman';font-size:16px">In the design of our biosensor, we used the part </span><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">BBa_J33201</span></span><span style=";font-family:'Times New Roman';font-size:16px"> registered By Edinburgh University in 2006 as the basis to build our biosensor, and installed a strong RBS and GFP reporter gene protein downstream of the part to build our biosensor.</span></p><p><span style=";font-family:'Times New Roman';font-size:16px"> </span></p><p><span style=";font-family:'Times New Roman';font-size:16px">The reason for this construction is that we got inspiration from the part </span><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">BBa_J33203</span></span><span style=";font-family:'Times New Roman';font-size:16px"> registered by the Edinburg Team in 2006, so we did not add terminator in the gene pathway when building this Biosensor.</span></p><p><span style=";font-family:'Times New Roman';font-size:16px"> </span></p><p><span style=";font-family:'Times New Roman';font-size:16px">Fortunately, we found that our biosensor could work normally and had a sensitive detection threshold for arsenic ions in water.</span></p><p><span style=";font-family:'Times New Roman';font-size:16px"> </span></p><p><span style=";font-family:'Times New Roman';font-size:16px">At the same time, we also characterized </span><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">J33201</span></span><span style=";font-family:'Times New Roman';font-size:16px">, an excellent part, and added more detailed and valuable data for the part.</span></p><p><span style=";font-family:'Times New Roman';font-size:16px">Our genetic pathway is designed as follows</span> </p><p style="text-align: center;"><img alt="Design(图1)" width="419" height="207" src="http://parts.igem.org/wiki/images/e/e4/T--Worldsher-wuhan-design_1.png" style="max-width:100%!important;height:auto;background:url(/public/plugins/Ueditor/lang/zh-cn/images/localimage.png) no-repeat center center;border:1px solid #ddd"><span style=";font-family:'Times New Roman';font-size:16px"> </span></p><p><span style=";font-family:'Times New Roman';font-size:16px"> </span></p><p><span style=";font-family:'Times New Roman';font-size:16px">J33201 is a part registered with the Edinburg Team in 2006, which includes an arsenic-sensitive promoter and </span><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">ArsR,</span></span><span style=";font-family:'Times New Roman';font-size:16px"> an auto-regulatory repressor from </span><em><span style="font-family: 'Times New Roman';font-size: 16px">E. coli.</span></em><span style=";font-family:'Times New Roman';font-size:16px"> On this basis, they constructed one of the biosensors, J3320 with a reporter gene of </span><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">lacZ</span></span><span style=";font-family:'Times New Roman';font-size:16px">, expressing galactosidase. </span></p><p><span style=";font-family:'Times New Roman';font-size:16px"> </span></p><p><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">LacZ</span></span><span style=";font-family:'Times New Roman';font-size:16px"> can react with substrate </span><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">x-gal</span></span><span style=";font-family:'Times New Roman';font-size:16px"> for colour development, yet we found in the preliminary experiment that this component was less sensitive to arsenic than expected, and the reaction time was very long.</span></p><p><span style=";font-family:'Times New Roman';font-size:16px"> </span></p><p><span style=";font-family:'Times New Roman';font-size:16px">So, we construct a biosensor expressing </span><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">GFP</span></span><span style=";font-family:'Times New Roman';font-size:16px">, and on this basis we characterize</span><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px"> J33201</span></span><span style=";font-family:'Times New Roman';font-size:16px"> again.</span></p><h1><strong><span style="font-family: 'Times New Roman';font-size: 16px">2.Biosorption</span></strong></h1><p><span style=";font-family:'Times New Roman';font-size:16px">Our biosorption consists of two components:</span></p><p><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">Metallothionein (MT)</span></span><span style=";font-family:'Times New Roman';font-size:16px"> plays an important role in bioremediation of heavy metals. </span><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">MT</span></span><span style=";font-family:'Times New Roman';font-size:16px"> is a kind of small molecular peptide rich in </span><em><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">cysteine (Cys-)</span></span></em><span style=";font-family:'Times New Roman';font-size:16px"> residues with low molecular weight. Its </span><em><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">Cys-</span></span></em><span style=";font-family:'Times New Roman';font-size:16px"> residues can adsorb many heavy metals such as lead, mercury and cadmium. However, since arsenic is a metalloid, these </span><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">MTs</span></span><span style=";font-family:'Times New Roman';font-size:16px"> have no specific adsorption on it. </span></p><p><span style=";font-family:'Times New Roman';font-size:16px"> </span></p><p><span style=";font-family:'Times New Roman';font-size:16px">Recently, scientists isolated and identified an </span><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">MT </span></span><span style=";font-family:'Times New Roman';font-size:16px">from a species of arsenic-tolerant Marine algae (</span><em><span style="font-family: 'Times New Roman';font-size: 16px">Fusus vesiculosus)</span></em><span style=";font-family:'Times New Roman';font-size:16px"> that, in vitro, demonstrated a high affinity for arsenic binding capacity.</span></p><p><span style=";font-family:'Times New Roman';font-size:16px"> </span></p><p><span style=";font-family:'Times New Roman';font-size:16px">In this project, we used the bacterial surface display technology to display </span><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">fMT</span></span><span style=";font-family:'Times New Roman';font-size:16px"> on the bacterial surface, and thus enhance the ability of absorbing arsenic.</span></p><p><span style=";font-family:'Times New Roman';font-size:16px"> </span></p><p><span style=";font-family:'Times New Roman';font-size:16px">Inspired by HUST-China team in 2014, the </span><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">fusion protein oprf-fMT</span></span><span style=";font-family:'Times New Roman';font-size:16px"> was constructed and heterogeneiously expressed by the </span><em><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">E. coli</span></span></em><span style=";font-family:'Times New Roman';font-size:16px"> expression system pET series plasmid.</span></p><p><span style=";font-family:'Times New Roman';font-size:16px"> </span></p><p><span style=";font-family:'Times New Roman';font-size:16px">Our design is as follows</span></p><p style="text-align: center;"><img alt="Design(图2)" width="273" height="163" src="http://parts.igem.org/wiki/images/7/72/T--Worldsher-wuhan-design_2.png" style="max-width:100%!important;height:auto;background:url(/public/plugins/Ueditor/lang/zh-cn/images/localimage.png) no-repeat center center;border:1px solid #ddd"><span style=";font-family:'Times New Roman';font-size:16px"> </span></p><p><span style=";font-family:'Times New Roman';font-size:16px"> </span></p><p style="text-align: center;"><img alt="Design(图3)" width="336" height="233" src="http://parts.igem.org/wiki/images/3/3b/T--Worldsher-wuhan-design_3.png" style="max-width:100%!important;height:auto;background:url(/public/plugins/Ueditor/lang/zh-cn/images/localimage.png) no-repeat center center;border:1px solid #ddd"><span style=";font-family:'Times New Roman';font-size:16px"> </span></p><p style="text-align: center;"><img alt="Design(图4)" width="449" height="198" src="http://parts.igem.org/wiki/images/9/9b/T-Worldsher-wuhan-design_4.png" style="max-width:100%!important;height:auto;background:url(/public/plugins/Ueditor/lang/zh-cn/images/localimage.png) no-repeat center center;border:1px solid #ddd"><span style=";font-family:'Times New Roman';font-size:16px"> </span></p><p><span style=";font-family:'Times New Roman';font-size:16px"> </span></p><p><span style=";font-family:'Times New Roman';font-size:16px">In addition, an important protein called </span><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">phytochelates (PC)</span></span><span style=";font-family:'Times New Roman';font-size:16px"> with a high affinity for arsenic has been found. PC is a natural peptide in plants and fungi, the general formula of the structure is</span><em><span style="font-family: 'Times New Roman';font-size: 16px"> </span></em><em><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">(</span></span></em><em><span style="text-decoration:underline;"><span style="font-family: 楷体;font-size: 16px">γ</span></span></em><em><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">-Glu-Cys-)n-Gly(n=1-11) </span></span></em><span style=";font-family:'Times New Roman';font-size:16px">. It was found that </span><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">PC</span></span><span style=";font-family:'Times New Roman';font-size:16px"> had a higher adsorption capacity for arsenic than </span><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">MT</span></span><span style=";font-family:'Times New Roman';font-size:16px">. </span><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">PC</span></span><span style=";font-family:'Times New Roman';font-size:16px"> is synthesized by </span><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">PC synthase (PCS)</span></span><span style=";font-family:'Times New Roman';font-size:16px"> with </span><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">glutathione (GSH)</span></span><span style=";font-family:'Times New Roman';font-size:16px"> as its substrate. </span></p><p><span style=";font-family:'Times New Roman';font-size:16px"> </span></p><p><span style=";font-family:'Times New Roman';font-size:16px">Previous studies have found that </span><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">PC synthase</span></span><span style=";font-family:'Times New Roman';font-size:16px"> from </span><em><span style="font-family: 'Times New Roman';font-size: 16px">Shizosomyces pombe</span></em><span style=";font-family:'Times New Roman';font-size:16px"> can be expressed in </span><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">SpPCS</span></span><span style=";font-family:'Times New Roman';font-size:16px"> in </span><em><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">E. coli</span></span></em><span style=";font-family:'Times New Roman';font-size:16px"> and thus enhance the adsorption ability of </span><em><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">E. coli</span></span></em><span style=";font-family:'Times New Roman';font-size:16px"> to arsenic.</span></p><p><span style=";font-family:'Times New Roman';font-size:16px"> </span></p><p><span style=";font-family:'Times New Roman';font-size:16px">Therefore, we artificially synthesized </span><span style="text-decoration:underline;"><span style="font-family: 'Times New Roman';font-size: 16px">ScPCS</span></span><span style=";font-family:'Times New Roman';font-size:16px"> genes (partial base changes to make them conform to RFC10 standard) and expressed them in pET expression system</span></p><p><span style=";font-family:'Times New Roman';font-size:16px"> </span></p><p><span style=";font-family:'Times New Roman';font-size:16px">Design is as follows</span></p><p style="text-align: center;"><img alt="Design(图5)" width="262" height="82" src="http://parts.igem.org/wiki/images/4/40/T-Worldsher-wuhan-design_5.png" style="max-width:100%!important;height:auto;background:url(/public/plugins/Ueditor/lang/zh-cn/images/localimage.png) no-repeat center center;border:1px solid #ddd"><span style=";font-family:楷体;font-size:16px"> </span></p><p><span style="text-decoration:underline;"><span style="font-family: 楷体;color: rgb(0, 0, 255);font-size: 16px"> </span></span></p><p style="text-align: center;"><img alt="Design(图6)" width="273" height="223" src="http://parts.igem.org/wiki/images/9/9f/T-Worldsher-wuhan-design_6.png" style="max-width:100%!important;height:auto;background:url(/public/plugins/Ueditor/lang/zh-cn/images/localimage.png) no-repeat center center;border:1px solid #ddd"><span style=";font-family:楷体;font-size:16px"> </span></p><p><span style=";font-family:楷体;font-size:16px"> </span></p><p style="text-align: center;"><img alt="Design(图7)" width="549" height="239" src="http://parts.igem.org/wiki/images/5/55/T-Worldsher-wuhan-design_7.png" style="max-width:100%!important;height:auto;background:url(/public/plugins/Ueditor/lang/zh-cn/images/localimage.png) no-repeat center center;border:1px solid #ddd"><span style=";font-family:楷体;font-size:16px"> </span></p></p><p><span style=";font-family:'Times New Roman';font-size:16px"> </span></p><p><span style=";font-family:'Times New Roman';font-size:16px">We hope that the synergistic effect can be achieved by constructing the recombinant bacteria with adsorption within the bacteria and the adsorption using surface display, and thus we can remove arsenic ions from water more effectively.</span></p><p><span style=";font-family:'Times New Roman';font-size:16px"> </span></p><p><span style=";font-family:'Times New Roman';font-size:16px">In conclusion, we intend to build a biosensor to monitor the arsenic concentration in water and use the bioremediation device we built to remove arsenic ions in water.</span></p><p><br></p> |
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