Difference between revisions of "Team:BGIC China"

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{{BGIC_China}}
 
{{BGIC_China}}
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Instead, we synthesized two relatively simple plamids(Fig.4 \u0026amp; Fig.5) with pCpxP promoter to test for the viability of the cell-free paper-based biochip system.\u003c/p\u003e","backupValue":"在此处添加文本段落。","version":1},{"type":"Image","id":"f_48a9ca92-51c6-4d30-b417-c0af4cb888ea","defaultValue":false,"link_url":"","thumb_url":"!","url":"!","caption":"","description":"","storageKey":"791914/屏幕截图_944__副本_ndu0g2","storage":"c","format":"png","h":1184,"w":1148,"s":126402,"new_target":true},{"type":"RichText","id":"f_b6a3e53c-8a5c-406c-8536-caedd740f884","defaultValue":false,"value":"\u003cp\u003e\u003cem\u003e\u003cstrong\u003eFig.4 pLV1C3-pCpxP-GFP\u003c/strong\u003e\u003c/em\u003e\u003c/p\u003e","backupValue":"在此处添加文本段落。","version":1},{"type":"Image","id":"f_83a4b6dd-a09b-4b4d-a9ab-ebd39c1a31ad","defaultValue":false,"link_url":"","thumb_url":"!","url":"!","caption":"","description":"","storageKey":"791914/屏幕截图_945__副本_ezzyeb","storage":"c","format":"png","h":1193,"w":1286,"s":134933,"new_target":true},{"type":"RichText","id":"f_f957f36b-fa8b-4893-9ac9-f2115dbc76b0","defaultValue":false,"value":"\u003cp\u003e\u003cem\u003e\u003cstrong\u003eFig.5 pLV1C3-pCpxP-LacZW\u003c/strong\u003e\u003c/em\u003e\u003c/p\u003e","backupValue":"在此处添加文本段落。","version":1},{"type":"RichText","id":"f_3a7cdbfa-984a-473c-80a1-afaccb6f2155","defaultValue":false,"value":"\u003cp style=\"font-size: 130%; text-align: center;\"\u003e\u003cem\u003epCpxP expression in cell\u003c/em\u003e\u003c/p\u003e\u003cp style=\"font-size:130%\"\u003eWe first transformed the plasmid pLV1C3-pCpxP-GFP into bacteria E.coli DH5α and test for the OD and RFU value under different concentrations of glucose and glycerol in order to characterize the pCpxP promoter. Based on a recent research, promoter CpxP was found to have potential in detecting a variaty of carbon sources in clinical samples[1]. However, our experimental results do not fit this research, the trend of our data generally cannot prove the inducibility of pCpxP by glucose or glycerol (Fig.6 \u0026amp; Fig.7).\u003c/p\u003e","backupValue":"在此处添加文本段落。","version":1},{"type":"Image","id":"f_762e583c-cd2c-43b6-a443-e8db68c7f4b8","defaultValue":false,"link_url":"","thumb_url":"!","url":"!","caption":"","description":"","storageKey":"791914/未命名_副本_dyroxs","storage":"c","format":"png","h":700,"w":1180,"s":8784,"new_target":true},{"type":"RichText","id":"f_bc11cb2f-01f1-4ae4-b28f-5249c65eb037","defaultValue":false,"value":"\u003cp\u003e\u003cem\u003e\u003cstrong\u003eFig.6 MRPU against glucose concentration (mg/dL)\u003c/strong\u003e\u003c/em\u003e\u003c/p\u003e","backupValue":"在此处添加文本段落。","version":1},{"type":"Image","id":"f_fb02a91c-fdcd-4b8c-a827-7d5a55f38b58","defaultValue":false,"link_url":"","thumb_url":"!","url":"!","caption":"","description":"","storageKey":"791914/未命名_副本2_vvg4os","storage":"c","format":"png","h":700,"w":1180,"s":8425,"new_target":true},{"type":"RichText","id":"f_e7c51c79-4337-47e2-835b-a91627302602","defaultValue":false,"value":"\u003cp\u003e\u003cem\u003e\u003cstrong\u003eFig.7 MRPU against glycerol concentration (mg/dL)\u003c/strong\u003e\u003c/em\u003e\u003c/p\u003e","backupValue":"在此处添加文本段落。","version":1},{"type":"RichText","id":"f_8edc8968-f2f3-4dc8-9a15-15dc0aef5334","defaultValue":false,"value":"\u003cp style=\"font-size: 130%;\"\u003eMRPU (modified relative promoter activity unit) is a equation that we modified from RPU[2] in order to determine the inducibility of a particular promoter by comparing the expression unit of the promoter with the expression unit of a reference promoter --- J23119 constitutive promoter. MRPU is a relatively simple equation that use much fewer parameters than RPU and provides a much easier way to perform the calculation to give a general idea about the inducibility of a particular promoter by particular factors.\u003c/p\u003e","backupValue":"在此处添加文本段落。","version":1},{"type":"Image","id":"f_61ee2497-b35b-4043-83f7-e961f2660a1a","defaultValue":false,"link_url":"","thumb_url":"!","url":"!","caption":"","description":"","storageKey":"791914/equations_rkq59o","storage":"c","format":"png","h":374,"w":618,"s":9309,"new_target":true},{"type":"RichText","id":"f_857c5be7-87cc-4311-91d6-5f5daab322d3","defaultValue":false,"value":"\u003cp\u003e\u003cem\u003e\u003cstrong\u003eFig.8 Equations used to calculate MRPU \u003c/strong\u003eHere we use pCpxP as promoter\u003c/em\u003e\u003c/p\u003e","backupValue":"在此处添加文本段落。","version":1},{"type":"RichText","id":"f_ed92cce4-51cc-40ed-906f-36bfef03350a","defaultValue":false,"value":"\u003cp style=\"font-size: 130%;\"\u003eAccording to Wolfe’s study, Cpx signal transduction system, in conjunction with SigmaE and Sigma32, responds to a board spectrum of adverse environmental conditions including high pH[3]. So we then test for the expression of promoter CpxP in E.coli DH5α in a range of pH from 6 to 9. We cultured bacteria in different pH under 37℃ for 10 hours and record the RFU and OD values. Graph of MRPU against pH has been plotted (Fig.9).\u003c/p\u003e","backupValue":"在此处添加文本段落。","version":1},{"type":"Image","id":"f_37a8eed5-8614-42da-839c-18246f561318","defaultValue":false,"link_url":"","thumb_url":"!","url":"!","caption":"","description":"","storageKey":"791914/graph_mumgfj","storage":"c","format":"png","h":809,"w":1274,"s":11850,"new_target":true},{"type":"RichText","id":"f_41f99526-ad7d-4e7d-8966-1498422db7b6","defaultValue":false,"value":"\u003cp\u003e\u003cem\u003e\u003cstrong\u003eFig.9 Graph of MRPU against pH\u003c/strong\u003e\u003c/em\u003e\u003c/p\u003e","backupValue":"在此处添加文本段落。","version":1},{"type":"RichText","id":"f_7fea12c4-8dc3-4fd0-b24c-2ec00cfcf560","defaultValue":false,"value":"\u003cp style=\"font-size: 130%;\"\u003eThe graph shows an logarithmic relationship between MRPU and pH, indicated the expression level of pCpxP increases when the pH rises. The MRPU at pH 9 is about three-folds the MRPU when pH is 6. Thus, the result shows that the expression level of pCpxP could be greatly increased when the pH has been increased in a certain range.\u003c/p\u003e","backupValue":"在此处添加文本段落。","version":1},{"type":"RichText","id":"f_26229fef-77fb-4340-9be9-7202436d96cb","defaultValue":false,"value":"\u003cp style=\"font-size:130%; text-align:center\"\u003e\u003cem\u003epCpxP expression in S30 cell-free system\u003c/em\u003e\u003c/p\u003e\u003cp style=\"font-size:130%\"\u003eNext, we put the pLV1C3-pCpxP-GFP and J23119-GFP plasmids into S30 cell-free system and record the RFU values of the mix after reaction for 1 hour to see if pCpxP promoter can express GFP protein in S30 cell-free system. The data proved that the pCpxP promoter can initiate the transcription in S30 cell-free system and has much higher expression level than J23119 promoter(Fig.10).\u003c/p\u003e","backupValue":"在此处添加文本段落。","version":1},{"type":"Image","id":"f_bfc2ac88-a9a7-479d-99ae-461ffe36ae6c","defaultValue":false,"link_url":"","thumb_url":"!","url":"!","caption":"","description":"","storageKey":"791914/graph2_icfbef","storage":"c","format":"png","h":746,"w":1256,"s":6570,"new_target":true},{"type":"RichText","id":"f_bb6f7cb1-5fd7-4323-a034-58919b018e21","defaultValue":false,"value":"\u003cp\u003e\u003cem\u003e\u003cstrong\u003eFig.10 Comparison between the RFU value of pCpxP-GFP and J23119-GFP plasmids in S30 cell-free system after cultured for 1 hour\u003c/strong\u003e\u003c/em\u003e\u003c/p\u003e","backupValue":"在此处添加文本段落。","version":1},{"type":"RichText","id":"f_010cc739-3544-48d4-846c-18ee7c1761c7","defaultValue":false,"value":"\u003cp style=\"font-size: 130%;\"\u003eDue to time issue, we are unable to test for the expression level of pCpxP in different pH in S30 cell-free system. 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</script> <!--[if lte IE 9]><script src="//assets.strikingly.com/assets/detectIE-b0fc7986f8eb7d40a31e9582bdeb7b88.js"></script><![endif]--> </head> <body class='show locale-zh-CN' id='user_sites'> <div id="s-page-container"><div data-reactroot="" data-reactid="1" data-react-checksum="-1555338099"><!-- react-empty: 2 --><div id="s-content" class="s-variation-default s-font-heading-default s-font-title-default s-font-body-default" data-reactid="3"><div class="social-thumbnail" data-reactid="4"><div style="display:none;" data-reactid="5"><img src="//assets.strikingly.com/assets/fb_images/default-715654470f8f9a657474bf68cefc382f.png" alt="page thumbnail" width="300" height="300" data-reactid="6"/></div><div id="lw-share-data" style="display:none;" data-reactid="7"><div class="lws-title" data-reactid="8">Ellz&#x27;s Site</div><div class="lws-description" data-reactid="9">Website</div><div class="lws-image" data-reactid="10">http://assets.strikingly.com/assets/fb_images/default-715654470f8f9a657474bf68cefc382f.png</div></div></div><!-- react-empty: 11 --><!-- react-empty: 12 --><ul class="slides s-page-1" data-reactid="13"><li class="slide s-section-1" id="section-f_3b3523a6-2671-4e7d-b310-e6dfeb26fc8d" data-reactid="14"><div class="waypoint" data-reactid="15"></div><a class="section-anchor" data-reactid="16"></a><div class="lazyload s-block-section s-section" data-reactid="17"><div class="container" data-reactid="18"><div class="columns sixteen" data-reactid="19"><!-- react-empty: 20 --></div><div class="alignment-container " data-reactid="21"><div class="s-repeatable s-block s-component s-mh " data-reactid="22"><div class="s-block-item s-repeatable-item s-block-sortable-item s-block-title " data-reactid="23"><span data-reactid="24"><div data-reactid="25"><div class="s-block-item-inner clearfix" style="position:relative;" data-reactid="26"><div class=" columns sixteen" data-reactid="27"><div class="s-item-title" data-reactid="28"><div class="s-component s-text" data-reactid="29"><h3 class="s-component-content s-font-heading" data-reactid="30"><p style="font-size: 130%;">The Proof of Concept</p></h3></div></div></div></div></div></span></div><div class="s-block-item s-repeatable-item s-block-sortable-item " data-reactid="31"><span data-reactid="32"><div data-reactid="33"><div class="s-block-item-inner clearfix" style="position:relative;" data-reactid="34"><div class=" columns sixteen" data-reactid="35"><div class="s-component s-text" data-reactid="36"><div class="s-component-content s-font-body" data-reactid="37"><p style="text-align: left; font-size: 130%;"><strong>Design:</strong></p><p style="text-align: left; font-size: 130%;"> </p><p style="text-align: left; font-size: 130%;">Our original design includes two plasmids, one acts as a sensor module (Fig.1), another acts as an signal amplification module (Fig.2).</p><p style="text-align: left; font-size: 130%;"> </p><p style="text-align: left; font-size: 130%;">The sensor plasmid contains pCpxP promoter which is inducible by various carbon sources including glucose, glycerol and citrate acid. Our experiment data also proved that the increase in pH could enchance the expression of pCpxP.</p></div></div></div></div></div></span></div><div class="s-block-item s-repeatable-item s-block-sortable-item " data-reactid="38"><span data-reactid="39"><div data-reactid="40"><div class="s-block-item-inner clearfix" style="position:relative;" data-reactid="41"><div class=" columns sixteen" data-reactid="42"><div class="s-component s-image" data-reactid="43"><span data-reactid="44"><div class="s-component-content" data-reactid="45"><div data-reactid="46"><!-- react-text: 47 --> <!-- /react-text --><div class="s-ratio-box" style="max-width:1267px;max-height:1193px;" data-reactid="48"><div class="s-ratio-fill" style="padding-bottom:94.16%;" data-reactid="49"></div><div data-reactid="50"><img alt="" title="" data-description="" width="1267" height="1193" src="data:image/gif;base64,R0lGODlhAQABAAAAACH5BAEKAAEALAAAAAABAAEAAAICTAEAOw==" class="lazyload " data-src="//res.cloudinary.com/hrscywv4p/image/upload/c_limit,h_9000,w_1200,f_auto,q_90/v1/791914/%E5%B1%8F%E5%B9%95%E6%88%AA%E5%9B%BE_940__%E5%89%AF%E6%9C%AC_vpa0bk.png" data-sizes="auto" data-reactid="51"/><noscript data-reactid="52"><img alt="" title="" data-description="" width="1267" height="1193" src="//res.cloudinary.com/hrscywv4p/image/upload/c_limit,h_9000,w_1200,f_auto,q_90/v1/791914/%E5%B1%8F%E5%B9%95%E6%88%AA%E5%9B%BE_940__%E5%89%AF%E6%9C%AC_vpa0bk.png" class="" data-reactid="53"/></noscript></div></div><!-- react-text: 54 --> <!-- /react-text --></div></div></span></div></div></div></div></span></div><div class="s-block-item s-repeatable-item s-block-sortable-item " data-reactid="55"><span data-reactid="56"><div data-reactid="57"><div class="s-block-item-inner clearfix" style="position:relative;" data-reactid="58"><div class=" columns sixteen" data-reactid="59"><div class="s-component s-text" data-reactid="60"><div class="s-component-content s-font-body" data-reactid="61"><p style="text-align: left;"><em><strong>Fig.1 Sensor plasmid (pLV1C3-pCpxP-LacZW-Cre)</strong></em></p></div></div></div></div></div></span></div><div class="s-block-item s-repeatable-item s-block-sortable-item " data-reactid="62"><span data-reactid="63"><div data-reactid="64"><div class="s-block-item-inner clearfix" style="position:relative;" data-reactid="65"><div class=" columns sixteen" data-reactid="66"><div class="s-component s-image" data-reactid="67"><span data-reactid="68"><div class="s-component-content" data-reactid="69"><div data-reactid="70"><!-- react-text: 71 --> <!-- /react-text --><div class="s-ratio-box" style="max-width:1281px;max-height:1191px;" data-reactid="72"><div class="s-ratio-fill" style="padding-bottom:92.97%;" data-reactid="73"></div><div data-reactid="74"><img alt="" title="" data-description="" width="1281" height="1191" src="data:image/gif;base64,R0lGODlhAQABAAAAACH5BAEKAAEALAAAAAABAAEAAAICTAEAOw==" class="lazyload " data-src="//res.cloudinary.com/hrscywv4p/image/upload/c_limit,h_9000,w_1200,f_auto,q_90/v1/791914/%E5%B1%8F%E5%B9%95%E6%88%AA%E5%9B%BE_941__%E5%89%AF%E6%9C%AC_faemrb.png" data-sizes="auto" data-reactid="75"/><noscript data-reactid="76"><img alt="" title="" data-description="" width="1281" height="1191" src="//res.cloudinary.com/hrscywv4p/image/upload/c_limit,h_9000,w_1200,f_auto,q_90/v1/791914/%E5%B1%8F%E5%B9%95%E6%88%AA%E5%9B%BE_941__%E5%89%AF%E6%9C%AC_faemrb.png" class="" data-reactid="77"/></noscript></div></div><!-- react-text: 78 --> <!-- /react-text --></div></div></span></div></div></div></div></span></div><div class="s-block-item s-repeatable-item s-block-sortable-item " data-reactid="79"><span data-reactid="80"><div data-reactid="81"><div class="s-block-item-inner clearfix" style="position:relative;" data-reactid="82"><div class=" columns sixteen" data-reactid="83"><div class="s-component s-text" data-reactid="84"><div class="s-component-content s-font-body" data-reactid="85"><p style="text-align: left;"><em><strong>Fig.2 Signal amplification plasmid (pET28a-LSL-LacZW-Cre)</strong></em></p></div></div></div></div></div></span></div><div class="s-block-item s-repeatable-item s-block-sortable-item " data-reactid="86"><span data-reactid="87"><div data-reactid="88"><div class="s-block-item-inner clearfix" style="position:relative;" data-reactid="89"><div class=" columns sixteen" data-reactid="90"><div class="s-component s-text" data-reactid="91"><div class="s-component-content s-font-body" data-reactid="92"><p style="text-align: left; font-size: 130%;">Sensor plasmid uses pCpxP promoter to control the expression level of the downstream genes---genes coding for LacZ and Cre. Cre is a recombinase which recognise loxp sites and recombine the target gene based on different directions of two loxp sites which flank the target gene. When Cre recombinase has been expressed from the sensor plasmid, it will cut down two B0015 terminators on the signal amplification plasmid and recombine the plasmid (Fig.3).</p></div></div></div></div></div></span></div><div class="s-block-item s-repeatable-item s-block-sortable-item " data-reactid="93"><span data-reactid="94"><div data-reactid="95"><div class="s-block-item-inner clearfix" style="position:relative;" data-reactid="96"><div class=" columns sixteen" data-reactid="97"><div class="s-component s-image" data-reactid="98"><span data-reactid="99"><div class="s-component-content" data-reactid="100"><div data-reactid="101"><!-- react-text: 102 --> <!-- /react-text --><div class="s-ratio-box" style="max-width:1962px;max-height:2073px;" data-reactid="103"><div class="s-ratio-fill" style="padding-bottom:105.66%;" data-reactid="104"></div><div data-reactid="105"><img alt="" title="" data-description="" width="1962" height="2073" src="data:image/gif;base64,R0lGODlhAQABAAAAACH5BAEKAAEALAAAAAABAAEAAAICTAEAOw==" class="lazyload " data-src="//res.cloudinary.com/hrscywv4p/image/upload/c_limit,h_9000,w_1200,f_auto,q_90/v1/791914/recombine_zszhla.png" data-sizes="auto" data-reactid="106"/><noscript data-reactid="107"><img alt="" title="" data-description="" width="1962" height="2073" src="//res.cloudinary.com/hrscywv4p/image/upload/c_limit,h_9000,w_1200,f_auto,q_90/v1/791914/recombine_zszhla.png" class="" data-reactid="108"/></noscript></div></div><!-- react-text: 109 --> <!-- /react-text --></div></div></span></div></div></div></div></span></div><div class="s-block-item s-repeatable-item s-block-sortable-item " data-reactid="110"><span data-reactid="111"><div data-reactid="112"><div class="s-block-item-inner clearfix" style="position:relative;" data-reactid="113"><div class=" columns sixteen" data-reactid="114"><div class="s-component s-text" data-reactid="115"><div class="s-component-content s-font-body" data-reactid="116"><p style="text-align: left;"><em><strong>Fig.3 Plasmid recombination by Cre at loxp sites</strong></em></p></div></div></div></div></div></span></div><div class="s-block-item s-repeatable-item s-block-sortable-item " data-reactid="117"><span data-reactid="118"><div data-reactid="119"><div class="s-block-item-inner clearfix" style="position:relative;" data-reactid="120"><div class=" columns sixteen" data-reactid="121"><div class="s-component s-text" data-reactid="122"><div class="s-component-content s-font-body" data-reactid="123"><p style="font-size: 130%;">When the B0015 terminators downstream of T7 promoter have been cut off, genes coding for LacZ and Cre could be expressed continuously given that IPTG is in excess supply. Chain reaction will be trigger because recombined signal amplification plasmid itself could express Cre recombinase that cut off B0015 terminators on intact signal amplification plasmids. Thus, the amount of LacZ will be accumulated in an exponential rate and the expression of LacZ will be sustained even pCpxP inducers are used up.</p></div></div></div></div></div></span></div><div class="s-block-item s-repeatable-item s-block-sortable-item " data-reactid="124"><span data-reactid="125"><div data-reactid="126"><div class="s-block-item-inner clearfix" style="position:relative;" data-reactid="127"><div class=" columns sixteen" data-reactid="128"><div class="s-component s-text" data-reactid="129"><div class="s-component-content s-font-body" data-reactid="130"><p style="font-size:130%"> </p><p style="font-size:130%"><strong>Experiment:</strong></p></div></div></div></div></div></span></div><div class="s-block-item s-repeatable-item s-block-sortable-item " data-reactid="131"><span data-reactid="132"><div data-reactid="133"><div class="s-block-item-inner clearfix" style="position:relative;" data-reactid="134"><div class=" columns sixteen" data-reactid="135"><div class="s-component s-text" data-reactid="136"><div class="s-component-content s-font-body" data-reactid="137"><p style="font-size: 130%; text-align: center;"><em>Plasmid assembly method</em></p><p style="font-size: 130%;">Genes coding for LacZ and Cre are obtained by PCR from plasmids which contain these two gene sequences with our designed primers. pCpxP promoter was synthesized by using phosphorylated single stranded oligonucleotides purchased from Invitrogen(TM). LSL(loxp-B0015-B0015-loxp) was also obtained by PCR with specially design primers. We then assembled our plasmids by using standard MCS subcloning method.</p></div></div></div></div></div></span></div><div class="s-block-item s-repeatable-item s-block-sortable-item " data-reactid="138"><span data-reactid="139"><div data-reactid="140"><div class="s-block-item-inner clearfix" style="position:relative;" data-reactid="141"><div class=" columns sixteen" data-reactid="142"><div class="s-component s-text" data-reactid="143"><div class="s-component-content s-font-body" data-reactid="144"><p style="font-size:130%; text-align:center"><em>Obstacles and changes of the original design</em></p><p style="font-size:130%">Due to technical and time issues, we are not able to follow our original design. Instead, we synthesized two relatively simple plamids(Fig.4 &amp; Fig.5) with pCpxP promoter to test for the viability of the cell-free paper-based biochip system.</p></div></div></div></div></div></span></div><div class="s-block-item s-repeatable-item s-block-sortable-item " data-reactid="145"><span data-reactid="146"><div data-reactid="147"><div class="s-block-item-inner clearfix" style="position:relative;" data-reactid="148"><div class=" columns sixteen" data-reactid="149"><div class="s-component s-image" data-reactid="150"><span data-reactid="151"><div class="s-component-content" data-reactid="152"><div data-reactid="153"><!-- react-text: 154 --> <!-- /react-text --><div class="s-ratio-box" style="max-width:1148px;max-height:1184px;" data-reactid="155"><div class="s-ratio-fill" style="padding-bottom:103.14%;" data-reactid="156"></div><div data-reactid="157"><img alt="" title="" data-description="" width="1148" height="1184" src="data:image/gif;base64,R0lGODlhAQABAAAAACH5BAEKAAEALAAAAAABAAEAAAICTAEAOw==" class="lazyload " data-src="//res.cloudinary.com/hrscywv4p/image/upload/c_limit,h_9000,w_1200,f_auto,q_90/v1/791914/%E5%B1%8F%E5%B9%95%E6%88%AA%E5%9B%BE_944__%E5%89%AF%E6%9C%AC_ndu0g2.png" data-sizes="auto" data-reactid="158"/><noscript data-reactid="159"><img alt="" title="" data-description="" width="1148" height="1184" src="//res.cloudinary.com/hrscywv4p/image/upload/c_limit,h_9000,w_1200,f_auto,q_90/v1/791914/%E5%B1%8F%E5%B9%95%E6%88%AA%E5%9B%BE_944__%E5%89%AF%E6%9C%AC_ndu0g2.png" class="" data-reactid="160"/></noscript></div></div><!-- react-text: 161 --> <!-- /react-text --></div></div></span></div></div></div></div></span></div><div class="s-block-item s-repeatable-item s-block-sortable-item " data-reactid="162"><span data-reactid="163"><div data-reactid="164"><div class="s-block-item-inner clearfix" style="position:relative;" data-reactid="165"><div class=" columns sixteen" data-reactid="166"><div class="s-component s-text" data-reactid="167"><div class="s-component-content s-font-body" data-reactid="168"><p><em><strong>Fig.4 pLV1C3-pCpxP-GFP</strong></em></p></div></div></div></div></div></span></div><div class="s-block-item s-repeatable-item s-block-sortable-item " data-reactid="169"><span data-reactid="170"><div data-reactid="171"><div class="s-block-item-inner clearfix" style="position:relative;" data-reactid="172"><div class=" columns sixteen" data-reactid="173"><div class="s-component s-image" data-reactid="174"><span data-reactid="175"><div class="s-component-content" data-reactid="176"><div data-reactid="177"><!-- react-text: 178 --> <!-- /react-text --><div class="s-ratio-box" style="max-width:1286px;max-height:1193px;" data-reactid="179"><div class="s-ratio-fill" style="padding-bottom:92.77%;" data-reactid="180"></div><div data-reactid="181"><img alt="" title="" data-description="" width="1286" height="1193" src="data:image/gif;base64,R0lGODlhAQABAAAAACH5BAEKAAEALAAAAAABAAEAAAICTAEAOw==" class="lazyload " data-src="//res.cloudinary.com/hrscywv4p/image/upload/c_limit,h_9000,w_1200,f_auto,q_90/v1/791914/%E5%B1%8F%E5%B9%95%E6%88%AA%E5%9B%BE_945__%E5%89%AF%E6%9C%AC_ezzyeb.png" data-sizes="auto" data-reactid="182"/><noscript data-reactid="183"><img alt="" title="" data-description="" width="1286" height="1193" src="//res.cloudinary.com/hrscywv4p/image/upload/c_limit,h_9000,w_1200,f_auto,q_90/v1/791914/%E5%B1%8F%E5%B9%95%E6%88%AA%E5%9B%BE_945__%E5%89%AF%E6%9C%AC_ezzyeb.png" class="" data-reactid="184"/></noscript></div></div><!-- react-text: 185 --> <!-- /react-text --></div></div></span></div></div></div></div></span></div><div class="s-block-item s-repeatable-item s-block-sortable-item " data-reactid="186"><span data-reactid="187"><div data-reactid="188"><div class="s-block-item-inner clearfix" style="position:relative;" data-reactid="189"><div class=" columns sixteen" data-reactid="190"><div class="s-component s-text" data-reactid="191"><div class="s-component-content s-font-body" data-reactid="192"><p><em><strong>Fig.5 pLV1C3-pCpxP-LacZW</strong></em></p></div></div></div></div></div></span></div><div class="s-block-item s-repeatable-item s-block-sortable-item " data-reactid="193"><span data-reactid="194"><div data-reactid="195"><div class="s-block-item-inner clearfix" style="position:relative;" data-reactid="196"><div class=" columns sixteen" data-reactid="197"><div class="s-component s-text" data-reactid="198"><div class="s-component-content s-font-body" data-reactid="199"><p style="font-size: 130%; text-align: center;"><em>pCpxP expression in cell</em></p><p style="font-size:130%">We first transformed the plasmid pLV1C3-pCpxP-GFP into bacteria E.coli DH5α and test for the OD and RFU value under different concentrations of glucose and glycerol in order to characterize the pCpxP promoter. Based on a recent research, promoter CpxP was found to have potential in detecting a variaty of carbon sources in clinical samples[1]. However, our experimental results do not fit this research, the trend of our data generally cannot prove the inducibility of pCpxP by glucose or glycerol (Fig.6 &amp; Fig.7).</p></div></div></div></div></div></span></div><div class="s-block-item s-repeatable-item s-block-sortable-item " data-reactid="200"><span data-reactid="201"><div data-reactid="202"><div class="s-block-item-inner clearfix" style="position:relative;" data-reactid="203"><div class=" columns sixteen" data-reactid="204"><div class="s-component s-image" data-reactid="205"><span data-reactid="206"><div class="s-component-content" data-reactid="207"><div data-reactid="208"><!-- react-text: 209 --> <!-- /react-text --><div class="s-ratio-box" style="max-width:1180px;max-height:700px;" data-reactid="210"><div class="s-ratio-fill" style="padding-bottom:59.32%;" data-reactid="211"></div><div data-reactid="212"><img alt="" title="" data-description="" width="1180" height="700" src="data:image/gif;base64,R0lGODlhAQABAAAAACH5BAEKAAEALAAAAAABAAEAAAICTAEAOw==" class="lazyload " data-src="//res.cloudinary.com/hrscywv4p/image/upload/c_limit,h_9000,w_1200,f_auto/v1/791914/%E6%9C%AA%E5%91%BD%E5%90%8D_%E5%89%AF%E6%9C%AC_dyroxs.png" data-sizes="auto" data-reactid="213"/><noscript data-reactid="214"><img alt="" title="" data-description="" width="1180" height="700" src="//res.cloudinary.com/hrscywv4p/image/upload/c_limit,h_9000,w_1200,f_auto/v1/791914/%E6%9C%AA%E5%91%BD%E5%90%8D_%E5%89%AF%E6%9C%AC_dyroxs.png" class="" data-reactid="215"/></noscript></div></div><!-- react-text: 216 --> <!-- /react-text --></div></div></span></div></div></div></div></span></div><div class="s-block-item s-repeatable-item s-block-sortable-item " data-reactid="217"><span data-reactid="218"><div data-reactid="219"><div class="s-block-item-inner clearfix" style="position:relative;" data-reactid="220"><div class=" columns sixteen" data-reactid="221"><div class="s-component s-text" data-reactid="222"><div class="s-component-content s-font-body" data-reactid="223"><p><em><strong>Fig.6 MRPU against glucose concentration (mg/dL)</strong></em></p></div></div></div></div></div></span></div><div class="s-block-item s-repeatable-item s-block-sortable-item " data-reactid="224"><span data-reactid="225"><div data-reactid="226"><div class="s-block-item-inner clearfix" style="position:relative;" data-reactid="227"><div class=" columns sixteen" data-reactid="228"><div class="s-component s-image" data-reactid="229"><span data-reactid="230"><div class="s-component-content" data-reactid="231"><div data-reactid="232"><!-- react-text: 233 --> <!-- /react-text --><div class="s-ratio-box" style="max-width:1180px;max-height:700px;" data-reactid="234"><div class="s-ratio-fill" style="padding-bottom:59.32%;" data-reactid="235"></div><div data-reactid="236"><img alt="" title="" data-description="" width="1180" height="700" src="data:image/gif;base64,R0lGODlhAQABAAAAACH5BAEKAAEALAAAAAABAAEAAAICTAEAOw==" class="lazyload " data-src="//res.cloudinary.com/hrscywv4p/image/upload/c_limit,h_9000,w_1200,f_auto/v1/791914/%E6%9C%AA%E5%91%BD%E5%90%8D_%E5%89%AF%E6%9C%AC2_vvg4os.png" data-sizes="auto" data-reactid="237"/><noscript data-reactid="238"><img alt="" title="" data-description="" width="1180" height="700" src="//res.cloudinary.com/hrscywv4p/image/upload/c_limit,h_9000,w_1200,f_auto/v1/791914/%E6%9C%AA%E5%91%BD%E5%90%8D_%E5%89%AF%E6%9C%AC2_vvg4os.png" class="" data-reactid="239"/></noscript></div></div><!-- react-text: 240 --> <!-- /react-text --></div></div></span></div></div></div></div></span></div><div class="s-block-item s-repeatable-item s-block-sortable-item " data-reactid="241"><span data-reactid="242"><div data-reactid="243"><div class="s-block-item-inner clearfix" style="position:relative;" data-reactid="244"><div class=" columns sixteen" data-reactid="245"><div class="s-component s-text" data-reactid="246"><div class="s-component-content s-font-body" data-reactid="247"><p><em><strong>Fig.7 MRPU against glycerol concentration (mg/dL)</strong></em></p></div></div></div></div></div></span></div><div class="s-block-item s-repeatable-item s-block-sortable-item " data-reactid="248"><span data-reactid="249"><div data-reactid="250"><div class="s-block-item-inner clearfix" style="position:relative;" data-reactid="251"><div class=" columns sixteen" data-reactid="252"><div class="s-component s-text" data-reactid="253"><div class="s-component-content s-font-body" data-reactid="254"><p style="font-size: 130%;">MRPU (modified relative promoter activity unit) is a equation that we modified from RPU[2] in order to determine the inducibility of a particular promoter by comparing the expression unit of the promoter with the expression unit of a reference promoter --- J23119 constitutive promoter. MRPU is a relatively simple equation that use much fewer parameters than RPU and provides a much easier way to perform the calculation to give a general idea about the inducibility of a particular promoter by particular factors.</p></div></div></div></div></div></span></div><div class="s-block-item s-repeatable-item s-block-sortable-item " data-reactid="255"><span data-reactid="256"><div data-reactid="257"><div class="s-block-item-inner clearfix" style="position:relative;" data-reactid="258"><div class=" columns sixteen" data-reactid="259"><div class="s-component s-image" data-reactid="260"><span data-reactid="261"><div class="s-component-content" data-reactid="262"><div data-reactid="263"><!-- react-text: 264 --> <!-- /react-text --><div class="s-ratio-box" style="max-width:618px;max-height:374px;" data-reactid="265"><div class="s-ratio-fill" style="padding-bottom:60.52%;" data-reactid="266"></div><div data-reactid="267"><img alt="" title="" data-description="" width="618" height="374" src="data:image/gif;base64,R0lGODlhAQABAAAAACH5BAEKAAEALAAAAAABAAEAAAICTAEAOw==" class="lazyload " data-src="//res.cloudinary.com/hrscywv4p/image/upload/c_limit,h_9000,w_1200,f_auto/v1/791914/equations_rkq59o.png" data-sizes="auto" data-reactid="268"/><noscript data-reactid="269"><img alt="" title="" data-description="" width="618" height="374" src="//res.cloudinary.com/hrscywv4p/image/upload/c_limit,h_9000,w_1200,f_auto/v1/791914/equations_rkq59o.png" class="" data-reactid="270"/></noscript></div></div><!-- react-text: 271 --> <!-- /react-text --></div></div></span></div></div></div></div></span></div><div class="s-block-item s-repeatable-item s-block-sortable-item " data-reactid="272"><span data-reactid="273"><div data-reactid="274"><div class="s-block-item-inner clearfix" style="position:relative;" data-reactid="275"><div class=" columns sixteen" data-reactid="276"><div class="s-component s-text" data-reactid="277"><div class="s-component-content s-font-body" data-reactid="278"><p><em><strong>Fig.8 Equations used to calculate MRPU </strong>Here we use pCpxP as promoter</em></p></div></div></div></div></div></span></div><div class="s-block-item s-repeatable-item s-block-sortable-item " data-reactid="279"><span data-reactid="280"><div data-reactid="281"><div class="s-block-item-inner clearfix" style="position:relative;" data-reactid="282"><div class=" columns sixteen" data-reactid="283"><div class="s-component s-text" data-reactid="284"><div class="s-component-content s-font-body" data-reactid="285"><p style="font-size: 130%;">According to Wolfe’s study, Cpx signal transduction system, in conjunction with SigmaE and Sigma32, responds to a board spectrum of adverse environmental conditions including high pH[3]. So we then test for the expression of promoter CpxP in E.coli DH5α in a range of pH from 6 to 9. We cultured bacteria in different pH under 37℃ for 10 hours and record the RFU and OD values. Graph of MRPU against pH has been plotted (Fig.9).</p></div></div></div></div></div></span></div><div class="s-block-item s-repeatable-item s-block-sortable-item " data-reactid="286"><span data-reactid="287"><div data-reactid="288"><div class="s-block-item-inner clearfix" style="position:relative;" data-reactid="289"><div class=" columns sixteen" data-reactid="290"><div class="s-component s-image" data-reactid="291"><span data-reactid="292"><div class="s-component-content" data-reactid="293"><div data-reactid="294"><!-- react-text: 295 --> <!-- /react-text --><div class="s-ratio-box" style="max-width:1274px;max-height:809px;" data-reactid="296"><div class="s-ratio-fill" style="padding-bottom:63.50%;" data-reactid="297"></div><div data-reactid="298"><img alt="" title="" data-description="" width="1274" height="809" src="data:image/gif;base64,R0lGODlhAQABAAAAACH5BAEKAAEALAAAAAABAAEAAAICTAEAOw==" class="lazyload " data-src="//res.cloudinary.com/hrscywv4p/image/upload/c_limit,h_9000,w_1200,f_auto/v1/791914/graph_mumgfj.png" data-sizes="auto" data-reactid="299"/><noscript data-reactid="300"><img alt="" title="" data-description="" width="1274" height="809" src="//res.cloudinary.com/hrscywv4p/image/upload/c_limit,h_9000,w_1200,f_auto/v1/791914/graph_mumgfj.png" class="" data-reactid="301"/></noscript></div></div><!-- react-text: 302 --> <!-- /react-text --></div></div></span></div></div></div></div></span></div><div class="s-block-item s-repeatable-item s-block-sortable-item " data-reactid="303"><span data-reactid="304"><div data-reactid="305"><div class="s-block-item-inner clearfix" style="position:relative;" data-reactid="306"><div class=" columns sixteen" data-reactid="307"><div class="s-component s-text" data-reactid="308"><div class="s-component-content s-font-body" data-reactid="309"><p><em><strong>Fig.9 Graph of MRPU against pH</strong></em></p></div></div></div></div></div></span></div><div class="s-block-item s-repeatable-item s-block-sortable-item " data-reactid="310"><span data-reactid="311"><div data-reactid="312"><div class="s-block-item-inner clearfix" style="position:relative;" data-reactid="313"><div class=" columns sixteen" data-reactid="314"><div class="s-component s-text" data-reactid="315"><div class="s-component-content s-font-body" data-reactid="316"><p style="font-size: 130%;">The graph shows an logarithmic relationship between MRPU and pH, indicated the expression level of pCpxP increases when the pH rises. The MRPU at pH 9 is about three-folds the MRPU when pH is 6. Thus, the result shows that the expression level of pCpxP could be greatly increased when the pH has been increased in a certain range.</p></div></div></div></div></div></span></div><div class="s-block-item s-repeatable-item s-block-sortable-item " data-reactid="317"><span data-reactid="318"><div data-reactid="319"><div class="s-block-item-inner clearfix" style="position:relative;" data-reactid="320"><div class=" columns sixteen" data-reactid="321"><div class="s-component s-text" data-reactid="322"><div class="s-component-content s-font-body" data-reactid="323"><p style="font-size:130%; text-align:center"><em>pCpxP expression in S30 cell-free system</em></p><p style="font-size:130%">Next, we put the pLV1C3-pCpxP-GFP and J23119-GFP plasmids into S30 cell-free system and record the RFU values of the mix after reaction for 1 hour to see if pCpxP promoter can express GFP protein in S30 cell-free system. The data proved that the pCpxP promoter can initiate the transcription in S30 cell-free system and has much higher expression level than J23119 promoter(Fig.10).</p></div></div></div></div></div></span></div><div class="s-block-item s-repeatable-item s-block-sortable-item " data-reactid="324"><span data-reactid="325"><div data-reactid="326"><div class="s-block-item-inner clearfix" style="position:relative;" data-reactid="327"><div class=" columns sixteen" data-reactid="328"><div class="s-component s-image" data-reactid="329"><span data-reactid="330"><div class="s-component-content" data-reactid="331"><div data-reactid="332"><!-- react-text: 333 --> <!-- /react-text --><div class="s-ratio-box" style="max-width:1256px;max-height:746px;" data-reactid="334"><div class="s-ratio-fill" style="padding-bottom:59.39%;" data-reactid="335"></div><div data-reactid="336"><img alt="" title="" data-description="" width="1256" height="746" src="data:image/gif;base64,R0lGODlhAQABAAAAACH5BAEKAAEALAAAAAABAAEAAAICTAEAOw==" class="lazyload " data-src="//res.cloudinary.com/hrscywv4p/image/upload/c_limit,h_9000,w_1200,f_auto/v1/791914/graph2_icfbef.png" data-sizes="auto" data-reactid="337"/><noscript data-reactid="338"><img alt="" title="" data-description="" width="1256" height="746" src="//res.cloudinary.com/hrscywv4p/image/upload/c_limit,h_9000,w_1200,f_auto/v1/791914/graph2_icfbef.png" class="" data-reactid="339"/></noscript></div></div><!-- react-text: 340 --> <!-- /react-text --></div></div></span></div></div></div></div></span></div><div class="s-block-item s-repeatable-item s-block-sortable-item " data-reactid="341"><span data-reactid="342"><div data-reactid="343"><div class="s-block-item-inner clearfix" style="position:relative;" data-reactid="344"><div class=" columns sixteen" data-reactid="345"><div class="s-component s-text" data-reactid="346"><div class="s-component-content s-font-body" data-reactid="347"><p><em><strong>Fig.10 Comparison between the RFU value of pCpxP-GFP and J23119-GFP plasmids in S30 cell-free system after cultured for 1 hour</strong></em></p></div></div></div></div></div></span></div><div class="s-block-item s-repeatable-item s-block-sortable-item " data-reactid="348"><span data-reactid="349"><div data-reactid="350"><div class="s-block-item-inner clearfix" style="position:relative;" data-reactid="351"><div class=" columns sixteen" data-reactid="352"><div class="s-component s-text" data-reactid="353"><div class="s-component-content s-font-body" data-reactid="354"><p style="font-size: 130%;">Due to time issue, we are unable to test for the expression level of pCpxP in different pH in S30 cell-free system. However, this data will be gained later and updated to our part pages.</p></div></div></div></div></div></span></div></div></div></div></div></li><li class="slide s-section-2 s-last-section-no-footer" id="section-f_cf699a11-f31b-493d-b50a-cbf5492a803d" data-reactid="355"><div class="waypoint" data-reactid="356"></div><a class="section-anchor" data-reactid="357"></a><div class="lazyload s-block-section s-section" data-reactid="358"><div class="container" data-reactid="359"><div class="columns sixteen" data-reactid="360"><!-- react-empty: 361 --></div><div class="alignment-container " data-reactid="362"><div class="s-repeatable s-block s-component s-mh " data-reactid="363"><div class="s-block-item s-repeatable-item s-block-sortable-item " data-reactid="364"><span data-reactid="365"><div data-reactid="366"><div class="s-block-item-inner clearfix" style="position:relative;" data-reactid="367"><div class=" columns sixteen" data-reactid="368"><div class="s-separator" data-reactid="369"><div class="s-separator-content" data-reactid="370"><div class="separator" data-reactid="371"></div></div></div></div></div></div></span></div><div class="s-block-item s-repeatable-item s-block-sortable-item " data-reactid="372"><span data-reactid="373"><div data-reactid="374"><div class="s-block-item-inner clearfix" style="position:relative;" data-reactid="375"><div class=" columns sixteen" data-reactid="376"><div class="s-component s-text" data-reactid="377"><div class="s-component-content s-font-body" data-reactid="378"><p style="font-size:130%; text-align:center"><strong><em>References</em></strong></p><p style="font-size:130%"> </p><p style="font-size:130%"><strong><em>1. Courbet, Alexis, et al. "Detection of pathological biomarkers in human clinical samples via amplifying genetic switches and logic gates." Science translational medicine 7.289 (2015): 289ra83-289ra83. </em></strong></p><p style="font-size:130%"><strong><em>2. Kelly, Jason R., et al. "Measuring the activity of BioBrick promoters using an in vivo reference standard." Journal of biological engineering 3.1 (2009): 1.​</em></strong></p><p style="font-size:130%"><em><strong>3. Wolfe, Alan J., et al. "Signal integration by the two-component signal transduction response regulator CpxR." Journal of bacteriology 190.7 (2008): 2314-2322.</strong></em></p></div></div></div></div></div></span></div></div></div></div></div><div class="waypoint" data-reactid="379"></div></li></ul><!-- react-empty: 380 --><div id="strikingly-tooltip-container" data-reactid="381"> </div><!-- react-empty: 382 --><!-- react-empty: 383 --><div class="s-floated-components" data-reactid="384"><div class="ecommerce-buy-dialog s-edit-modal extend" id="ecommerce-buy-dialog" data-reactid="385"><div class="s-async-wrapper s-component" data-reactid="386"><div class="s-loading" data-reactid="387"></div></div></div><div id="s-ecommerce-shopping-cart-wrapper" class="s-ecommerce-shopping-cart-wrapper" data-reactid="388"></div></div></div></div></div> <div id='fb-root'></div> <div id='app-script-root'></div> <div id='app-view-root'></div> <script id='blog-collection-tmpl' type='text/html'> <div class='s-blog-col-head'></div> <div class='s-blog-col-body'> {{ _.each(blogPosts, function(blog) { }} <div class='s-blog-entry'> <div class='s-blog-entry-inner'> {{ if ($S && $S.conf && $S.conf.preview_mode) { }} <a class='s-blog-overlay-link' href='{{=blog.publicUrl}}'>&nbsp;</a> {{ } else { }} <a class='s-blog-overlay-link' href='{{=blog.relativeUrl}}'>&nbsp;</a> {{ } }} {{ if (blog.iconUrl) { }} <div class='s-blog-entry-left'> {{ if (UrlHelper.imageHasContent(blog.iconUrl)) { }} <div class='s-blog-avatar' style='background-image:url({{=blog.iconUrl}});'></div> {{ } else { }} <div class='s-blog-default-avatar'> <div class='entypo-bookmark'></div> </div> {{ } }} </div> {{ } }} <div class='s-blog-entry-right'> <div class='s-blog-details'> <div class='s-blog-details-head s-blog-row'> <div class='s-blog-title s-font-heading'> {{=blog.title}} </div> <div class='s-blog-date'> {{=blog.publishedAt}} </div> </div> {{ if (blogSettings.previewLayout == 1) { }} <div class='s-blog-details-blurb s-blog-row'> {{=blog.blurb}} </div> {{ } }} </div> </div> </div> </div> {{ }) }} {{ if (blogPosts.length == 0) { }} <div class='s-blog-entry'> <div class='s-common-status no-posts-error'> 没有博文。 </div> </div> {{ } }} </div> {{ if (blogPosts.length > 0) { }} <div class='s-blog-col-foot'> <a class='s-blog-link s-blog-next-link' href='javascript:void(0);'> &laquo; 更新 </a> &nbsp; <ul class='s-blog-pagination' style='display:none;'> <li> <a class='s-blog-link s-blog-pagination-page' href='javascript:void(0);'></a> </li> </ul> &nbsp; <a class='s-blog-link s-blog-prev-link' href='javascript:void(0);'> 其它 &raquo; </a> </div> {{ } }} </script> <script src="//cdn.bootcss.com/jquery/1.10.0/jquery.min.js"></script> <script>
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</style><div class="s-navbar-section" data-reactid="12"><div id="header-container" class="navigator " data-reactid="13"><div class="nav-left" data-reactid="14"><span class="nav-items" data-reactid="15"></span></div><div class="nav-center" data-reactid="16"><div class="logo" data-reactid="17"><div class="s-component s-image" data-reactid="18"><span data-reactid="19"><div class="s-component-content" data-reactid="20"><div data-reactid="21"><!-- react-text: 22 --> <!-- /react-text --><img src="//res.cloudinary.com/hrscywv4p/image/upload/c_limit,h_1440,w_720,f_auto,q_90/v1/791914/Team_logo_final_%E5%89%AF%E6%9C%AC_%E5%89%AF%E6%9C%AC_ffweyb.png" alt="" title="" data-description="" data-reactid="23"/><!-- react-text: 24 --> <!-- /react-text --></div></div></span></div></div></div><div class="nav-right" data-reactid="25"><span class="nav-items" data-reactid="26"></span></div><div class="nav original-nav" data-reactid="27"><span data-reactid="28"><span class="nav-item" style="display:none;" data-reactid="29"><a class="s-nav-item" href="#mx48" data-reactid="30"><span class="s-font-body" data-reactid="31">Title</span></a></span><span class="nav-item" style="display:none;" data-reactid="32"><a class="s-nav-item" href="#biografia" data-reactid="33"><span class="s-font-body" data-reactid="34">Slide</span></a></span><span class="nav-item" style="display:none;" data-reactid="35"><a class="s-nav-item" href="#conocenos" data-reactid="36"><span class="s-font-body" data-reactid="37">Project Abstract</span></a></span><span class="nav-item" style="display:none;" data-reactid="38"><a class="s-nav-item" href="#blog48" data-reactid="39"><span class="s-font-body" data-reactid="40">Other parts</span></a></span></span><span data-reactid="41"><li data-reactid="42"><a class="s-external-link-item s-nav-item" href="https://2016.igem.org/Team:BGIC_China/Description" target="_blank" data-reactid="43"><span class="s-font-body" data-reactid="44">Project description</span></a></li><li data-reactid="45"><a class="s-external-link-item s-nav-item" href="https://2016.igem.org/Team:BGIC_China/Proof" target="_blank" data-reactid="46"><span class="s-font-body" data-reactid="47">Proof of concept</span></a></li><li data-reactid="48"><a class="s-external-link-item s-nav-item" href="https://2016.igem.org/Team:BGIC_China/Parts" target="_blank" data-reactid="49"><span class="s-font-body" data-reactid="50">Parts</span></a></li><li data-reactid="51"><a class="s-external-link-item s-nav-item" href="https://2016.igem.org/Team:BGIC_China/HP/Gold" target="_blank" data-reactid="52"><span class="s-font-body" data-reactid="53">Human practice</span></a></li><li data-reactid="54"><a class="s-external-link-item s-nav-item" href="https://2016.igem.org/Team:BGIC_China/Attributions" target="_blank" data-reactid="55"><span class="s-font-body" data-reactid="56">Attributions</span></a></li><li data-reactid="57"><a class="s-external-link-item s-nav-item" href="https://2016.igem.org/Team:BGIC_China/Collaborations" target="_blank" data-reactid="58"><span class="s-font-body" data-reactid="59">Collaborations</span></a></li></span></div></div><div id="header-container-fixed" class="navigator strikingly-fixed" data-reactid="60"><div class="nav-left" data-reactid="61"><span class="nav-items" data-reactid="62"></span></div><div class="nav-center" data-reactid="63"><div class="logo" data-reactid="64"><div class="s-component s-image" data-reactid="65"><span data-reactid="66"><div class="s-component-content" data-reactid="67"><div data-reactid="68"><!-- react-text: 69 --> <!-- /react-text --><img src="//res.cloudinary.com/hrscywv4p/image/upload/c_limit,h_1440,w_720,f_auto,q_90/v1/791914/Team_logo_final_%E5%89%AF%E6%9C%AC_%E5%89%AF%E6%9C%AC_ffweyb.png" alt="" title="" data-description="" data-reactid="70"/><!-- react-text: 71 --> <!-- /react-text --></div></div></span></div></div></div><div class="nav-right" data-reactid="72"><span class="nav-items" data-reactid="73"></span></div></div></div><!-- react-empty: 74 --><ul class="slides s-page-1" data-reactid="75"><li class="slide s-section-1" id="section-f_2689c0d2-ada7-41e9-97eb-0be2cf91b15f" data-reactid="76"><div class="waypoint" data-reactid="77"></div><a class="section-anchor" data-reactid="78"></a><div class="s-bg-image s-bg-light-text s-bg-overlay s-cta-section s-section" style="background-repeat:no-repeat;background-size:cover;background-color:transparent;background-position:50% 50%;background-image:url(//res.cloudinary.com/hrscywv4p/image/upload/c_limit,h_1500,w_2000,f_auto,q_90/v1/791914/pic1_mntejh.jpg);" data-reactid="79"><div class="container s-layout-mediaTop" data-reactid="80"><div class="columns sixteen" data-reactid="81"><div class="s-title-group " data-reactid="82"><div class="s-title " data-reactid="83"><div class="s-component s-text" data-reactid="84"><h1 class="s-component-content s-font-title" data-reactid="85"><p style="font-size:80%" class="s-text-color-custom2">Paper, but more than a paper</p></h1></div></div><div class="s-subtitle" data-reactid="86"><div class="s-component s-text" data-reactid="87"><h4 class="s-component-content s-font-heading" data-reactid="88"><p class="s-text-color-custom2">Programmable and open-source Next Generation Bio-Chip (NGBC) based on cell-free system</p></h4></div></div></div><div class="s-button-group " data-reactid="89"><div class="s-button s-component" data-reactid="90"><span data-reactid="91"><div class="s-component-content" data-reactid="92"><a class="s-action-button s-common-button s-font-body" href="#2" data-component="button" target="_self" data-reactid="93">READ MORE ↓</a></div></span></div></div></div></div></div></li><li class="slide s-section-2" id="section-f_16951574-e142-4b54-86c5-19cba5f9cb49" data-reactid="94"><div class="waypoint" data-reactid="95"></div><a class="section-anchor" data-reactid="96"></a><div class="s-section s-slider-section" data-reactid="97"><div class="s-component s-slider " data-reactid="98"><!-- react-text: 99 --> <!-- /react-text --><!-- react-text: 100 --> <!-- /react-text --><div class="s-component-content" data-reactid="101"><div class="slider-container loading" data-reactid="102"><div data-auto-play="3500" class="iosslider" data-reactid="103"><div class="slider" data-reactid="104"><div class="lazyload s-bg-image s-bg-light-text s-bg-overlay item" style="background-repeat:repeat;background-size:auto;background-color:transparent;background-position:50% 50%;background-image:url(data:image/gif;base64,R0lGODlhAQABAAAAACH5BAEKAAEALAAAAAABAAEAAAICTAEAOw==);" data-bg="//uploads.strikinglycdn.com/static/backgrounds/pastel-solids/49.jpg" data-reactid="105"><div class="inner" data-reactid="106"><div class="container right s-rva" data-reactid="107"><div class="eight columns right media-outer s-rva-media" data-reactid="108"><div class="media-wrapper" data-reactid="109"><div class="s-component s-media " data-reactid="110"><span data-reactid="111"><div class="s-component-content" data-reactid="112"><div data-reactid="113"><!-- react-text: 114 --> <!-- /react-text --><div class="s-ratio-box" style="max-width:200px;max-height:200px;" data-reactid="115"><div class="s-ratio-fill" style="padding-bottom:100.00%;" data-reactid="116"></div><div data-reactid="117"><img alt="" title="" data-description="" width="200" height="200" src="data:image/gif;base64,R0lGODlhAQABAAAAACH5BAEKAAEALAAAAAABAAEAAAICTAEAOw==" class="lazyload " data-src="//res.cloudinary.com/hrscywv4p/image/upload/c_limit,h_1440,w_720,f_auto/v1/791914/programmable_200_pajmy7.png" data-sizes="auto" data-reactid="118"/><noscript data-reactid="119"><img alt="" title="" data-description="" width="200" height="200" src="//res.cloudinary.com/hrscywv4p/image/upload/c_limit,h_1440,w_720,f_auto/v1/791914/programmable_200_pajmy7.png" class="" data-reactid="120"/></noscript></div></div><!-- react-text: 121 --> <!-- /react-text --></div></div></span></div></div></div><div class="columns right left-align valign slider-desc s-rva-text eight" data-reactid="122"><div class="half-offset-right" data-reactid="123"><div class="s-title-group " data-reactid="124"><div class="s-title " data-reactid="125"><div class="s-component s-text" data-reactid="126"><h2 class="s-component-content s-font-title" data-reactid="127"><p class="s-text-color-custom2">Programmable</p></h2></div></div><div class="s-subtitle" data-reactid="128"><div class="s-component s-text" data-reactid="129"><h4 class="s-component-content s-font-heading" data-reactid="130"><p class="s-text-color-custom2">Give bio-chip different functions by re-construct immobilized genetic circuits.</p></h4></div></div></div><div class="cta-button-wrapper" data-reactid="131"><div class="" data-reactid="132"><div class="s-button s-component" data-reactid="133"><span data-reactid="134"><div class="s-component-content" data-reactid="135"></div></span></div></div></div></div></div></div></div></div><div class="lazyload s-bg-image s-bg-light-text s-bg-overlay item" style="background-repeat:repeat;background-size:auto;background-color:transparent;background-position:50% 50%;background-image:url(data:image/gif;base64,R0lGODlhAQABAAAAACH5BAEKAAEALAAAAAABAAEAAAICTAEAOw==);" data-bg="//uploads.strikinglycdn.com/static/backgrounds/pastel-solids/49.jpg" data-reactid="136"><div class="inner" data-reactid="137"><div class="container right s-rva" data-reactid="138"><div class="eight columns right media-outer s-rva-media" data-reactid="139"><div class="media-wrapper" data-reactid="140"><div class="s-component s-media " data-reactid="141"><span data-reactid="142"><div class="s-component-content" data-reactid="143"><div data-reactid="144"><!-- react-text: 145 --> <!-- /react-text --><div class="s-ratio-box" style="max-width:200px;max-height:200px;" data-reactid="146"><div class="s-ratio-fill" style="padding-bottom:100.00%;" data-reactid="147"></div><div data-reactid="148"><img alt="" title="" data-description="" width="200" height="200" src="data:image/gif;base64,R0lGODlhAQABAAAAACH5BAEKAAEALAAAAAABAAEAAAICTAEAOw==" class="lazyload " data-src="//res.cloudinary.com/hrscywv4p/image/upload/c_limit,h_1440,w_720,f_auto/v1/791914/Cell-free_iia73w.png" data-sizes="auto" data-reactid="149"/><noscript data-reactid="150"><img alt="" title="" data-description="" width="200" height="200" src="//res.cloudinary.com/hrscywv4p/image/upload/c_limit,h_1440,w_720,f_auto/v1/791914/Cell-free_iia73w.png" class="" data-reactid="151"/></noscript></div></div><!-- react-text: 152 --> <!-- /react-text --></div></div></span></div></div></div><div class="columns right left-align valign slider-desc s-rva-text eight" data-reactid="153"><div class="half-offset-right" data-reactid="154"><div class="s-title-group " data-reactid="155"><div class="s-title " data-reactid="156"><div class="s-component s-text" data-reactid="157"><h2 class="s-component-content s-font-title" data-reactid="158"><p class="s-text-color-custom2">Cell-Free</p></h2></div></div><div class="s-subtitle" data-reactid="159"><div class="s-component s-text" data-reactid="160"><h4 class="s-component-content s-font-heading" data-reactid="161"><p class="s-text-color-custom2">Bio-safety, low noise and more possibilities.</p></h4></div></div></div><div class="cta-button-wrapper" data-reactid="162"><div class="" data-reactid="163"><div class="s-button s-component" data-reactid="164"><span data-reactid="165"><div class="s-component-content" data-reactid="166"></div></span></div></div></div></div></div></div></div></div><div class="lazyload s-bg-image s-bg-light-text s-bg-overlay item" style="background-repeat:repeat;background-size:auto;background-color:transparent;background-position:50% 50%;background-image:url(data:image/gif;base64,R0lGODlhAQABAAAAACH5BAEKAAEALAAAAAABAAEAAAICTAEAOw==);" data-bg="//uploads.strikinglycdn.com/static/backgrounds/pastel-solids/49.jpg" data-reactid="167"><div class="inner" data-reactid="168"><div class="container right s-rva" data-reactid="169"><div class="eight columns right media-outer s-rva-media" data-reactid="170"><div class="media-wrapper" data-reactid="171"><div class="s-component s-media " data-reactid="172"><span data-reactid="173"><div class="s-component-content" data-reactid="174"><div data-reactid="175"><!-- react-text: 176 --> <!-- /react-text --><div class="s-ratio-box" style="max-width:200px;max-height:200px;" data-reactid="177"><div class="s-ratio-fill" style="padding-bottom:100.00%;" data-reactid="178"></div><div data-reactid="179"><img alt="" title="" data-description="" width="200" height="200" src="data:image/gif;base64,R0lGODlhAQABAAAAACH5BAEKAAEALAAAAAABAAEAAAICTAEAOw==" class="lazyload " data-src="//res.cloudinary.com/hrscywv4p/image/upload/c_limit,h_1440,w_720,f_auto/v1/791914/open_source_woezqa.png" data-sizes="auto" data-reactid="180"/><noscript data-reactid="181"><img alt="" title="" data-description="" width="200" height="200" src="//res.cloudinary.com/hrscywv4p/image/upload/c_limit,h_1440,w_720,f_auto/v1/791914/open_source_woezqa.png" class="" data-reactid="182"/></noscript></div></div><!-- react-text: 183 --> <!-- /react-text --></div></div></span></div></div></div><div class="columns right left-align valign slider-desc s-rva-text eight" data-reactid="184"><div class="half-offset-right" data-reactid="185"><div class="s-title-group " data-reactid="186"><div class="s-title " data-reactid="187"><div class="s-component s-text" data-reactid="188"><h2 class="s-component-content s-font-title" data-reactid="189"><p class="s-text-color-custom2">Open-Source</p></h2></div></div><div class="s-subtitle" data-reactid="190"><div class="s-component s-text" data-reactid="191"><h4 class="s-component-content s-font-heading" data-reactid="192"><p class="s-text-color-custom2">Distribute technology around the world.</p></h4></div></div></div><div class="cta-button-wrapper" data-reactid="193"><div class="" data-reactid="194"><div class="s-button s-component" data-reactid="195"><span data-reactid="196"><div class="s-component-content" data-reactid="197"></div></span></div></div></div></div></div></div></div></div><div class="lazyload s-bg-image s-bg-light-text s-bg-overlay item" style="background-repeat:repeat;background-size:auto;background-color:transparent;background-position:50% 50%;background-image:url(data:image/gif;base64,R0lGODlhAQABAAAAACH5BAEKAAEALAAAAAABAAEAAAICTAEAOw==);" data-bg="//uploads.strikinglycdn.com/static/backgrounds/pastel-solids/49.jpg" data-reactid="198"><div class="inner" data-reactid="199"><div class="container right s-rva" data-reactid="200"><div class="eight columns right media-outer s-rva-media" data-reactid="201"><div class="media-wrapper" data-reactid="202"><div class="s-component s-media " data-reactid="203"><span data-reactid="204"><div class="s-component-content" data-reactid="205"><div data-reactid="206"><!-- react-text: 207 --> <!-- /react-text --><div class="s-ratio-box" style="max-width:200px;max-height:200px;" data-reactid="208"><div class="s-ratio-fill" style="padding-bottom:100.00%;" data-reactid="209"></div><div data-reactid="210"><img alt="" title="" data-description="" width="200" height="200" src="data:image/gif;base64,R0lGODlhAQABAAAAACH5BAEKAAEALAAAAAABAAEAAAICTAEAOw==" class="lazyload " data-src="//res.cloudinary.com/hrscywv4p/image/upload/c_limit,h_1440,w_720,f_auto/v1/791914/short_prototyping_time_ijzly8.png" data-sizes="auto" data-reactid="211"/><noscript data-reactid="212"><img alt="" title="" data-description="" width="200" height="200" src="//res.cloudinary.com/hrscywv4p/image/upload/c_limit,h_1440,w_720,f_auto/v1/791914/short_prototyping_time_ijzly8.png" class="" data-reactid="213"/></noscript></div></div><!-- react-text: 214 --> <!-- /react-text --></div></div></span></div></div></div><div class="columns right left-align valign slider-desc s-rva-text eight" data-reactid="215"><div class="half-offset-right" data-reactid="216"><div class="s-title-group " data-reactid="217"><div class="s-title " data-reactid="218"><div class="s-component s-text" data-reactid="219"><h2 class="s-component-content s-font-title" data-reactid="220"><p class="s-text-color-custom2">Short Prototyping Time</p></h2></div></div><div class="s-subtitle" data-reactid="221"><div class="s-component s-text" data-reactid="222"><h4 class="s-component-content s-font-heading" data-reactid="223"><p class="s-text-color-custom2">Build a new bio-chip just with in a week.</p></h4></div></div></div><div class="cta-button-wrapper" data-reactid="224"><div class="" data-reactid="225"><div class="s-button s-component" data-reactid="226"><span data-reactid="227"><div class="s-component-content" data-reactid="228"></div></span></div></div></div></div></div></div></div></div><div class="lazyload s-bg-image s-bg-light-text s-bg-overlay item" style="background-repeat:repeat;background-size:auto;background-color:transparent;background-position:50% 50%;background-image:url(data:image/gif;base64,R0lGODlhAQABAAAAACH5BAEKAAEALAAAAAABAAEAAAICTAEAOw==);" data-bg="//uploads.strikinglycdn.com/static/backgrounds/pastel-solids/49.jpg" data-reactid="229"><div class="inner" data-reactid="230"><div class="container right s-rva" data-reactid="231"><div class="eight columns right media-outer s-rva-media" data-reactid="232"><div class="media-wrapper" data-reactid="233"><div class="s-component s-media " data-reactid="234"><span data-reactid="235"><div class="s-component-content" data-reactid="236"><div data-reactid="237"><!-- react-text: 238 --> <!-- /react-text --><div class="s-ratio-box" style="max-width:200px;max-height:200px;" data-reactid="239"><div class="s-ratio-fill" style="padding-bottom:100.00%;" data-reactid="240"></div><div data-reactid="241"><img alt="" title="" data-description="" width="200" height="200" src="data:image/gif;base64,R0lGODlhAQABAAAAACH5BAEKAAEALAAAAAABAAEAAAICTAEAOw==" class="lazyload " data-src="//res.cloudinary.com/hrscywv4p/image/upload/c_limit,h_1440,w_720,f_auto/v1/791914/963306978_%E5%89%AF%E6%9C%AC_oththa.png" data-sizes="auto" data-reactid="242"/><noscript data-reactid="243"><img alt="" title="" data-description="" width="200" height="200" src="//res.cloudinary.com/hrscywv4p/image/upload/c_limit,h_1440,w_720,f_auto/v1/791914/963306978_%E5%89%AF%E6%9C%AC_oththa.png" class="" data-reactid="244"/></noscript></div></div><!-- react-text: 245 --> <!-- /react-text --></div></div></span></div></div></div><div class="columns right left-align valign slider-desc s-rva-text eight" data-reactid="246"><div class="half-offset-right" data-reactid="247"><div class="s-title-group " data-reactid="248"><div class="s-title " data-reactid="249"><div class="s-component s-text" data-reactid="250"><h2 class="s-component-content s-font-title" data-reactid="251"><p class="s-text-color-custom2">One-Step &amp; High-Throughput</p></h2></div></div><div class="s-subtitle" data-reactid="252"><div class="s-component s-text" data-reactid="253"><h4 class="s-component-content s-font-heading" data-reactid="254"><p class="s-text-color-custom2">Complete all the reactions on one bio-chip and test for multiple samples together.</p></h4></div></div></div><div class="cta-button-wrapper" data-reactid="255"><div class="" data-reactid="256"><div class="s-button s-component" data-reactid="257"><span data-reactid="258"><div class="s-component-content" data-reactid="259"></div></span></div></div></div></div></div></div></div></div><div class="lazyload s-bg-image s-bg-light-text s-bg-overlay item" style="background-repeat:repeat;background-size:auto;background-color:transparent;background-position:50% 50%;background-image:url(data:image/gif;base64,R0lGODlhAQABAAAAACH5BAEKAAEALAAAAAABAAEAAAICTAEAOw==);" data-bg="//uploads.strikinglycdn.com/static/backgrounds/pastel-solids/49.jpg" data-reactid="260"><div class="inner" data-reactid="261"><div class="container right s-rva" data-reactid="262"><div class="eight columns right media-outer s-rva-media" data-reactid="263"><div class="media-wrapper" data-reactid="264"><div class="s-component s-media " data-reactid="265"><span data-reactid="266"><div class="s-component-content" data-reactid="267"><div data-reactid="268"><!-- react-text: 269 --> <!-- /react-text --><div class="s-ratio-box" style="max-width:200px;max-height:200px;" data-reactid="270"><div class="s-ratio-fill" style="padding-bottom:100.00%;" data-reactid="271"></div><div data-reactid="272"><img alt="" title="" data-description="" width="200" height="200" src="data:image/gif;base64,R0lGODlhAQABAAAAACH5BAEKAAEALAAAAAABAAEAAAICTAEAOw==" class="lazyload " data-src="//res.cloudinary.com/hrscywv4p/image/upload/c_limit,h_1440,w_720,f_auto/v1/791914/Ultra-high_expression_htz7ij.png" data-sizes="auto" data-reactid="273"/><noscript data-reactid="274"><img alt="" title="" data-description="" width="200" height="200" src="//res.cloudinary.com/hrscywv4p/image/upload/c_limit,h_1440,w_720,f_auto/v1/791914/Ultra-high_expression_htz7ij.png" class="" data-reactid="275"/></noscript></div></div><!-- react-text: 276 --> <!-- /react-text --></div></div></span></div></div></div><div class="columns right left-align valign slider-desc s-rva-text eight" data-reactid="277"><div class="half-offset-right" data-reactid="278"><div class="s-title-group " data-reactid="279"><div class="s-title " data-reactid="280"><div class="s-component s-text" data-reactid="281"><h2 class="s-component-content s-font-title" data-reactid="282"><p class="s-text-color-custom2">Ultra-High Expression</p></h2></div></div><div class="s-subtitle" data-reactid="283"><div class="s-component s-text" data-reactid="284"><h4 class="s-component-content s-font-heading" data-reactid="285"><p class="s-text-color-custom2">U-HEC (Utra-High Expression Cassette) enables high protein expression on the bio-chip.</p></h4></div></div></div><div class="cta-button-wrapper" data-reactid="286"><div class="" data-reactid="287"><div class="s-button s-component" data-reactid="288"><span data-reactid="289"><div class="s-component-content" data-reactid="290"></div></span></div></div></div></div></div></div></div></div><div class="lazyload s-bg-image s-bg-light-text s-bg-overlay item" style="background-repeat:repeat;background-size:auto;background-color:transparent;background-position:50% 50%;background-image:url(data:image/gif;base64,R0lGODlhAQABAAAAACH5BAEKAAEALAAAAAABAAEAAAICTAEAOw==);" data-bg="//uploads.strikinglycdn.com/static/backgrounds/pastel-solids/49.jpg" data-reactid="291"><div class="inner" data-reactid="292"><div class="container right s-rva" data-reactid="293"><div class="eight columns right media-outer s-rva-media" data-reactid="294"><div class="media-wrapper" data-reactid="295"><div class="s-component s-media " data-reactid="296"><span data-reactid="297"><div class="s-component-content" data-reactid="298"><div data-reactid="299"><!-- react-text: 300 --> <!-- /react-text --><div class="s-ratio-box" style="max-width:200px;max-height:200px;" data-reactid="301"><div class="s-ratio-fill" style="padding-bottom:100.00%;" data-reactid="302"></div><div data-reactid="303"><img alt="" title="" data-description="" width="200" height="200" src="data:image/gif;base64,R0lGODlhAQABAAAAACH5BAEKAAEALAAAAAABAAEAAAICTAEAOw==" class="lazyload " data-src="//res.cloudinary.com/hrscywv4p/image/upload/c_limit,h_1440,w_720,f_auto/v1/791914/unlimited_potentials_eqmpdt.png" data-sizes="auto" data-reactid="304"/><noscript data-reactid="305"><img alt="" title="" data-description="" width="200" height="200" src="//res.cloudinary.com/hrscywv4p/image/upload/c_limit,h_1440,w_720,f_auto/v1/791914/unlimited_potentials_eqmpdt.png" class="" data-reactid="306"/></noscript></div></div><!-- react-text: 307 --> <!-- /react-text --></div></div></span></div></div></div><div class="columns right left-align valign slider-desc s-rva-text eight" data-reactid="308"><div class="half-offset-right" data-reactid="309"><div class="s-title-group " data-reactid="310"><div class="s-title " data-reactid="311"><div class="s-component s-text" data-reactid="312"><h2 class="s-component-content s-font-title" data-reactid="313"><p class="s-text-color-custom2">Unlimited Potentials</p></h2></div></div><div class="s-subtitle" data-reactid="314"><div class="s-component s-text" data-reactid="315"><h4 class="s-component-content s-font-heading" data-reactid="316"><p class="s-text-color-custom2">It is possible to testing for everything by construct proper genetic circuits.</p></h4></div></div></div><div class="cta-button-wrapper" data-reactid="317"><div class="" data-reactid="318"><div class="s-button s-component" data-reactid="319"><span data-reactid="320"><div class="s-component-content" data-reactid="321"></div></span></div></div></div></div></div></div></div></div></div><div class="center slide-selectors" data-reactid="322"><div class="slide-selectors-inner" data-reactid="323"><div class="content repeatable-component" data-reactid="324"><div class="selector-wrapper" data-reactid="325"><div class="selector" data-reactid="326"></div></div><div class="selector-wrapper" data-reactid="327"><div class="selector" data-reactid="328"></div></div><div class="selector-wrapper" data-reactid="329"><div class="selector" data-reactid="330"></div></div><div class="selector-wrapper" data-reactid="331"><div class="selector" data-reactid="332"></div></div><div class="selector-wrapper" data-reactid="333"><div class="selector" data-reactid="334"></div></div><div class="selector-wrapper" data-reactid="335"><div class="selector" data-reactid="336"></div></div><div class="selector-wrapper" data-reactid="337"><div class="selector" data-reactid="338"></div></div><div class="clearfix" data-reactid="339"></div></div></div></div><div class="prev-button" data-reactid="340"><div class="arrow" data-reactid="341"></div></div><div class="next-button" data-reactid="342"><div class="arrow" data-reactid="343"></div></div></div></div></div></div></div></li><li class="slide s-section-3" id="section-f_498d3f75-a829-4591-b39c-655949bf5eda" data-reactid="344"><div class="waypoint" data-reactid="345"></div><a class="section-anchor" data-reactid="346"></a><div class="lazyload s-no-bg s-block-section s-section" data-reactid="347"><div class="container" data-reactid="348"><div class="columns sixteen" data-reactid="349"><div class="s-title-group " data-reactid="350"><div class="s-title " data-reactid="351"><div class="s-component s-text" data-reactid="352"><h2 class="s-component-content s-font-title" data-reactid="353"><p>Project Abstract</p></h2></div></div></div></div><div class="alignment-container " data-reactid="354"><div class="s-repeatable s-block s-component s-mh " data-reactid="355"><div class="s-block-item s-repeatable-item s-block-sortable-item " data-reactid="356"><span data-reactid="357"><div data-reactid="358"><div class="s-block-item-inner clearfix" style="position:relative;" data-reactid="359"><div class=" columns sixteen" data-reactid="360"><div class="s-component s-text" data-reactid="361"><div class="s-component-content s-font-body" data-reactid="362"><p style="font-size:130%; text-align:left">NGBC --- Next Generation Bio-Chip, is what we are going to create. This will be the first bio-chip ever that allows user with knowledge in synthetic biology to easily re-program and re-construct it in order to make new bio-chips with different functions.</p></div></div></div></div></div></span></div><div class="s-block-item s-repeatable-item s-block-sortable-item " data-reactid="363"><span data-reactid="364"><div data-reactid="365"><div class="s-block-item-inner clearfix" style="position:relative;" data-reactid="366"><div class=" columns sixteen" data-reactid="367"><div class="s-component s-text" data-reactid="368"><div class="s-component-content s-font-body" data-reactid="369"><p style="font-size:130%; text-align:left">NGBC use genetic circuits to process the input signals and given output results. In order to minimize the disruptions and limitations of biochemical reactions inside a living-cell, the reaction center of NGBC will use S30 cell-free system which is produced from cell extracts and could provide all the conditions required for biochemical reactions like that inside a real living-cell. What is most amazing is that the complete genetic circuits and cell-free system is just carried by simple filter paper and no other expensive materials required to construct it. We are also developing a hardware and a software which can be integrated with NGBC to provide a more accurate data readout and a more interactive user environment.</p></div></div></div></div></div></span></div><div class="s-block-item s-repeatable-item s-block-sortable-item " data-reactid="370"><span data-reactid="371"><div data-reactid="372"><div class="s-block-item-inner clearfix" style="position:relative;" data-reactid="373"><div class=" columns sixteen" data-reactid="374"><div class="s-component s-text" data-reactid="375"><div class="s-component-content s-font-body" data-reactid="376"><p style="font-size:130%; text-align:left">In future, we intended to develop a completely open-source NGBC kit which contains all the necessary materials, protocols and genetic components required to re-program the BioChip, and the kit will be distributed to labs around the world so there will be more new BioChips with different functions be developed. We will also create an NGBC clinical database that includes all the verified BioChips which are able to measure the concentration of target clinical molecules such as glucose, nitrate and fatty-acids in the clinical sample with in a certain range.</p></div></div></div></div></div></span></div></div></div></div></div></li><li class="slide s-section-4 s-last-section-no-footer" id="section-f_dc10117f-1cc6-41a0-9381-17b786992ba0" data-reactid="377"><div class="waypoint" data-reactid="378"></div><a class="section-anchor" data-reactid="379"></a><div class="lazyload s-bg-image s-bg-light-text s-block-section s-section" style="background-repeat:repeat;background-size:auto;background-color:transparent;background-position:50% 50%;background-image:url(data:image/gif;base64,R0lGODlhAQABAAAAACH5BAEKAAEALAAAAAABAAEAAAICTAEAOw==);" data-bg="//uploads.strikinglycdn.com/static/backgrounds/pastel-solids/34.jpg" data-reactid="380"><div class="container" data-reactid="381"><div class="columns sixteen" data-reactid="382"><!-- react-empty: 383 --></div><div class="alignment-container " data-reactid="384"><div class="s-repeatable s-block s-component s-mh " data-reactid="385"><div class="s-block-item s-repeatable-item s-block-sortable-item s-block-smallFeatureLeft " data-reactid="386"><span data-reactid="387"><div data-reactid="388"><div class="s-block-item-inner clearfix" style="position:relative;" data-reactid="389"><div class="" data-reactid="390"><div class="s-block-feature " data-reactid="391"><div class="six columns" data-reactid="392"><div class="s-item-media-group" data-reactid="393"><div class="s-component s-media " data-reactid="394"><span data-reactid="395"><div class="s-component-content" data-reactid="396"><div data-reactid="397"><a href="https://2016.igem.org/Team:BGIC_China/Description" target="_blank" data-reactid="398"><div class="s-ratio-box" style="max-width:160px;max-height:160px;" data-reactid="399"><div class="s-ratio-fill" style="padding-bottom:100.00%;" data-reactid="400"></div><div data-reactid="401"><img alt="" title="" data-description="" width="160" height="160" src="data:image/gif;base64,R0lGODlhAQABAAAAACH5BAEKAAEALAAAAAABAAEAAAICTAEAOw==" class="lazyload " data-src="//uploads.strikinglycdn.com/static/icons/line-icons/8.png" data-sizes="auto" data-reactid="402"/><noscript data-reactid="403"><img alt="" title="" data-description="" width="160" height="160" src="//uploads.strikinglycdn.com/static/icons/line-icons/8.png" class="" data-reactid="404"/></noscript></div></div></a><!-- react-text: 405 --> <!-- /react-text --><!-- react-text: 406 --> <!-- /react-text --></div></div></span></div></div></div><div class="ten columns" data-reactid="407"><div class="s-item-text-group half-offset-left" data-reactid="408"><div class="s-item-title" data-reactid="409"><div class="s-component s-text" data-reactid="410"><h3 class="s-component-content s-font-heading" data-reactid="411"><p><a target="_blank" href="https://2016.igem.org/Team:BGIC_China/Description"><strong>Project description</strong></a></p></h3></div></div><div class="s-item-subtitle" data-reactid="412"><div class="s-component s-text" data-reactid="413"><h6 class="s-component-content s-font-body" data-reactid="414"><p>We are going to design the first programmable BioChip ever by using genetic boolean logic gates(Piro et al, 2013) which enable the BioChip to achieve complex tasks. Several other amazing features like cell-free expression system and ultra-high expression cassette(U-HEC) are also introduced...</p></h6></div></div><div class="s-item-text" data-reactid="415"><div class="s-component s-text" data-reactid="416"><div class="s-component-content s-font-body" data-reactid="417"><p><a target="_blank" href="https://2016.igem.org/Team:BGIC_China/Description"><strong>READ MORE</strong></a></p></div></div></div></div></div></div></div></div></div></span></div><div class="s-block-item s-repeatable-item s-block-sortable-item s-block-smallFeatureRight " data-reactid="418"><span data-reactid="419"><div data-reactid="420"><div class="s-block-item-inner clearfix" style="position:relative;" data-reactid="421"><div class="" data-reactid="422"><div class="s-block-feature " data-reactid="423"><div class="six columns right" data-reactid="424"><div class="s-item-media-group" data-reactid="425"><div class="s-component s-media " data-reactid="426"><span data-reactid="427"><div class="s-component-content" data-reactid="428"><div data-reactid="429"><a href="https://2016.igem.org/Team:BGIC_China/Proof" target="_blank" data-reactid="430"><div class="s-ratio-box" style="max-width:160px;max-height:160px;" data-reactid="431"><div class="s-ratio-fill" style="padding-bottom:100.00%;" data-reactid="432"></div><div data-reactid="433"><img alt="" title="" data-description="" width="160" height="160" src="data:image/gif;base64,R0lGODlhAQABAAAAACH5BAEKAAEALAAAAAABAAEAAAICTAEAOw==" class="lazyload " data-src="//uploads.strikinglycdn.com/static/icons/line-icons/49.png" data-sizes="auto" data-reactid="434"/><noscript data-reactid="435"><img alt="" title="" data-description="" width="160" height="160" src="//uploads.strikinglycdn.com/static/icons/line-icons/49.png" class="" data-reactid="436"/></noscript></div></div></a><!-- react-text: 437 --> <!-- /react-text --><!-- react-text: 438 --> <!-- /react-text --></div></div></span></div></div></div><div class="ten columns right" data-reactid="439"><div class="s-item-text-group half-offset-right" data-reactid="440"><div class="s-item-title" data-reactid="441"><div class="s-component s-text" data-reactid="442"><h3 class="s-component-content s-font-heading" data-reactid="443"><p><a target="_blank" href="https://2016.igem.org/Team:BGIC_China/Proof"><strong>Proof of concept</strong></a></p></h3></div></div><div class="s-item-subtitle" data-reactid="444"><div class="s-component s-text" data-reactid="445"><h6 class="s-component-content s-font-body" data-reactid="446"><p>In order to prove the feasibility of our idea, we constructed a simple plasmid with a pH inducible promoter with EGFP coding sequence in the downstream of the promoter. We immobilized this plasmid onto our bio-chip system and tested for the RFU value in comparison with a standard constitutive J23119 promoter ...</p></h6></div></div><div class="s-item-text" data-reactid="447"><div class="s-component s-text" data-reactid="448"><div class="s-component-content s-font-body" data-reactid="449"><p><a target="_blank" href="https://2016.igem.org/Team:BGIC_China/Proof"><strong>READ MORE</strong></a></p></div></div></div></div></div></div></div></div></div></span></div><div class="s-block-item s-repeatable-item s-block-sortable-item s-block-smallFeatureLeft " data-reactid="450"><span data-reactid="451"><div data-reactid="452"><div class="s-block-item-inner clearfix" style="position:relative;" data-reactid="453"><div class="" data-reactid="454"><div class="s-block-feature " data-reactid="455"><div class="six columns" data-reactid="456"><div class="s-item-media-group" data-reactid="457"><div class="s-component s-media " data-reactid="458"><span data-reactid="459"><div class="s-component-content" data-reactid="460"><div data-reactid="461"><a href="https://2016.igem.org/Team:BGIC_China/Parts" target="_blank" data-reactid="462"><div class="s-ratio-box" style="max-width:160px;max-height:160px;" data-reactid="463"><div class="s-ratio-fill" style="padding-bottom:100.00%;" data-reactid="464"></div><div data-reactid="465"><img alt="" title="" data-description="" width="160" height="160" src="data:image/gif;base64,R0lGODlhAQABAAAAACH5BAEKAAEALAAAAAABAAEAAAICTAEAOw==" class="lazyload " data-src="//uploads.strikinglycdn.com/static/icons/line-icons/33.png" data-sizes="auto" data-reactid="466"/><noscript data-reactid="467"><img alt="" title="" data-description="" width="160" height="160" src="//uploads.strikinglycdn.com/static/icons/line-icons/33.png" class="" data-reactid="468"/></noscript></div></div></a><!-- react-text: 469 --> <!-- /react-text --><!-- react-text: 470 --> <!-- /react-text --></div></div></span></div></div></div><div class="ten columns" data-reactid="471"><div class="s-item-text-group half-offset-left" data-reactid="472"><div class="s-item-title" data-reactid="473"><div class="s-component s-text" data-reactid="474"><h3 class="s-component-content s-font-heading" data-reactid="475"><p><a target="_blank" href="https://2016.igem.org/Team:BGIC_China/Parts"><strong>Parts</strong></a></p></h3></div></div><div class="s-item-subtitle" data-reactid="476"><div class="s-component s-text" data-reactid="477"><h6 class="s-component-content s-font-body" data-reactid="478"><p>BBa_K2026000 --- a composite part with two B0015 sequence flanked by two same directional loxp sites.</p><p>BBa_K2026001 --- pH and glucose inducible ultra-strong promoter from Cpx regulon.</p></h6></div></div><div class="s-item-text" data-reactid="479"><div class="s-component s-text" data-reactid="480"><div class="s-component-content s-font-body" data-reactid="481"><p><a target="_blank" href="https://2016.igem.org/Team:BGIC_China/Parts"><strong>READ MORE</strong></a></p></div></div></div></div></div></div></div></div></div></span></div><div class="s-block-item s-repeatable-item s-block-sortable-item s-block-smallFeatureRight " data-reactid="482"><span data-reactid="483"><div data-reactid="484"><div class="s-block-item-inner clearfix" style="position:relative;" data-reactid="485"><div class="" data-reactid="486"><div class="s-block-feature " data-reactid="487"><div class="six columns right" data-reactid="488"><div class="s-item-media-group" data-reactid="489"><div class="s-component s-media " data-reactid="490"><span data-reactid="491"><div class="s-component-content" data-reactid="492"><div data-reactid="493"><a href="https://2016.igem.org/Team:BGIC_China/HP/Gold" target="_blank" data-reactid="494"><div class="s-ratio-box" style="max-width:160px;max-height:160px;" data-reactid="495"><div class="s-ratio-fill" style="padding-bottom:100.00%;" data-reactid="496"></div><div data-reactid="497"><img alt="" title="" data-description="" width="160" height="160" src="data:image/gif;base64,R0lGODlhAQABAAAAACH5BAEKAAEALAAAAAABAAEAAAICTAEAOw==" class="lazyload " data-src="//uploads.strikinglycdn.com/static/icons/line-icons/15.png" data-sizes="auto" data-reactid="498"/><noscript data-reactid="499"><img alt="" title="" data-description="" width="160" height="160" src="//uploads.strikinglycdn.com/static/icons/line-icons/15.png" class="" data-reactid="500"/></noscript></div></div></a><!-- react-text: 501 --> <!-- /react-text --><!-- react-text: 502 --> <!-- /react-text --></div></div></span></div></div></div><div class="ten columns right" data-reactid="503"><div class="s-item-text-group half-offset-right" data-reactid="504"><div class="s-item-title" data-reactid="505"><div class="s-component s-text" data-reactid="506"><h3 class="s-component-content s-font-heading" data-reactid="507"><p><a target="_blank" href="https://2016.igem.org/Team:BGIC_China/HP/Gold"><strong>Human practice</strong></a></p></h3></div></div><div class="s-item-subtitle" data-reactid="508"><div class="s-component s-text" data-reactid="509"><h6 class="s-component-content s-font-body" data-reactid="510"><p>We introduced our project to visitors in London International Youth Science Forum (LIYSF) and spread the concept of synthetic biology including bio-safety to high school students and Bio-Maker organisations...</p></h6></div></div><div class="s-item-text" data-reactid="511"><div class="s-component s-text" data-reactid="512"><div class="s-component-content s-font-body" data-reactid="513"><p><a target="_blank" href="https://2016.igem.org/Team:BGIC_China/HP/Gold"><strong>READ MORE</strong></a></p></div></div></div></div></div></div></div></div></div></span></div><div class="s-block-item s-repeatable-item s-block-sortable-item s-block-smallFeatureLeft " data-reactid="514"><span data-reactid="515"><div data-reactid="516"><div class="s-block-item-inner clearfix" style="position:relative;" data-reactid="517"><div class="" data-reactid="518"><div class="s-block-feature " data-reactid="519"><div class="six columns" data-reactid="520"><div class="s-item-media-group" data-reactid="521"><div class="s-component s-media " data-reactid="522"><span data-reactid="523"><div class="s-component-content" data-reactid="524"><div data-reactid="525"><a href="https://2016.igem.org/Team:BGIC_China/Attributions" target="_blank" data-reactid="526"><div class="s-ratio-box" style="max-width:160px;max-height:160px;" data-reactid="527"><div class="s-ratio-fill" style="padding-bottom:100.00%;" data-reactid="528"></div><div data-reactid="529"><img alt="" title="" data-description="" width="160" height="160" src="data:image/gif;base64,R0lGODlhAQABAAAAACH5BAEKAAEALAAAAAABAAEAAAICTAEAOw==" class="lazyload " data-src="//uploads.strikinglycdn.com/static/icons/line-icons/93.png" data-sizes="auto" data-reactid="530"/><noscript data-reactid="531"><img alt="" title="" data-description="" width="160" height="160" src="//uploads.strikinglycdn.com/static/icons/line-icons/93.png" class="" data-reactid="532"/></noscript></div></div></a><!-- react-text: 533 --> <!-- /react-text --><!-- react-text: 534 --> <!-- /react-text --></div></div></span></div></div></div><div class="ten columns" data-reactid="535"><div class="s-item-text-group half-offset-left" data-reactid="536"><div class="s-item-title" data-reactid="537"><div class="s-component s-text" data-reactid="538"><h3 class="s-component-content s-font-heading" data-reactid="539"><p><a target="_blank" href="https://2016.igem.org/Team:BGIC_China/Attributions"><strong>Attributions</strong></a></p></h3></div></div><div class="s-item-subtitle" data-reactid="540"><div class="s-component s-text" data-reactid="541"><h6 class="s-component-content s-font-body" data-reactid="542"><p>Thanks to all the team members including our instructors and advisers who make great contributes to the project. We are also flattered to introduce our sponsors and donators...</p></h6></div></div><div class="s-item-text" data-reactid="543"><div class="s-component s-text" data-reactid="544"><div class="s-component-content s-font-body" data-reactid="545"><p><a target="_blank" href="https://2016.igem.org/Team:BGIC_China/Attributions"><strong>READ MORE</strong></a></p></div></div></div></div></div></div></div></div></div></span></div><div class="s-block-item s-repeatable-item s-block-sortable-item s-block-smallFeatureRight " data-reactid="546"><span data-reactid="547"><div data-reactid="548"><div class="s-block-item-inner clearfix" style="position:relative;" data-reactid="549"><div class="" data-reactid="550"><div class="s-block-feature " data-reactid="551"><div class="six columns right" data-reactid="552"><div class="s-item-media-group" data-reactid="553"><div class="s-component s-media " data-reactid="554"><span data-reactid="555"><div class="s-component-content" data-reactid="556"><div data-reactid="557"><a href="https://2016.igem.org/Team:BGIC_China/Collaborations" target="_blank" data-reactid="558"><div class="s-ratio-box" style="max-width:160px;max-height:160px;" data-reactid="559"><div class="s-ratio-fill" style="padding-bottom:100.00%;" data-reactid="560"></div><div data-reactid="561"><img alt="" title="" data-description="" width="160" height="160" src="data:image/gif;base64,R0lGODlhAQABAAAAACH5BAEKAAEALAAAAAABAAEAAAICTAEAOw==" class="lazyload " data-src="//uploads.strikinglycdn.com/static/icons/line-icons/34.png" data-sizes="auto" data-reactid="562"/><noscript data-reactid="563"><img alt="" title="" data-description="" width="160" height="160" src="//uploads.strikinglycdn.com/static/icons/line-icons/34.png" class="" data-reactid="564"/></noscript></div></div></a><!-- react-text: 565 --> <!-- /react-text --><!-- react-text: 566 --> <!-- /react-text --></div></div></span></div></div></div><div class="ten columns right" data-reactid="567"><div class="s-item-text-group half-offset-right" data-reactid="568"><div class="s-item-title" data-reactid="569"><div class="s-component s-text" data-reactid="570"><h3 class="s-component-content s-font-heading" data-reactid="571"><p><a target="_blank" href="https://2016.igem.org/Team:BGIC_China/Collaborations"><strong>Collaborations</strong></a></p></h3></div></div><div class="s-item-subtitle" data-reactid="572"><div class="s-component s-text" data-reactid="573"></div></div><div class="s-item-text" data-reactid="574"><div class="s-component s-text" data-reactid="575"><div class="s-component-content s-font-body" data-reactid="576"><p><a target="_blank" href="https://2016.igem.org/Team:BGIC_China/Collaborations"><strong>READ MORE</strong></a></p></div></div></div></div></div></div></div></div></div></span></div></div></div></div></div><div class="waypoint" data-reactid="577"></div></li></ul><!-- react-empty: 578 --><div id="strikingly-tooltip-container" data-reactid="579"> </div><!-- react-empty: 580 --><!-- react-empty: 581 --><div class="s-floated-components" data-reactid="582"><div class="ecommerce-buy-dialog s-edit-modal extend" id="ecommerce-buy-dialog" data-reactid="583"><div class="s-async-wrapper s-component" data-reactid="584"><div class="s-loading" data-reactid="585"></div></div></div><div id="s-ecommerce-shopping-cart-wrapper" class="s-ecommerce-shopping-cart-wrapper" data-reactid="586"></div></div></div></div></div> <div id='fb-root'></div> <div id='app-script-root'></div> <div id='app-view-root'></div> <script id='blog-collection-tmpl' type='text/html'> <div class='s-blog-col-head'></div> <div class='s-blog-col-body'> {{ _.each(blogPosts, function(blog) { }} <div class='s-blog-entry'> <div class='s-blog-entry-inner'> {{ if ($S && $S.conf && $S.conf.preview_mode) { }} <a class='s-blog-overlay-link' href='{{=blog.publicUrl}}'>&nbsp;</a> {{ } else { }} <a class='s-blog-overlay-link' href='{{=blog.relativeUrl}}'>&nbsp;</a> {{ } }} {{ if (blog.iconUrl) { }} <div class='s-blog-entry-left'> {{ if (UrlHelper.imageHasContent(blog.iconUrl)) { }} <div class='s-blog-avatar' style='background-image:url({{=blog.iconUrl}});'></div> {{ } else { }} <div class='s-blog-default-avatar'> <div class='entypo-bookmark'></div> </div> {{ } }} </div> {{ } }} <div class='s-blog-entry-right'> <div class='s-blog-details'> <div class='s-blog-details-head s-blog-row'> <div class='s-blog-title s-font-heading'> {{=blog.title}} </div> <div class='s-blog-date'> {{=blog.publishedAt}} </div> </div> {{ if (blogSettings.previewLayout == 1) { }} <div class='s-blog-details-blurb s-blog-row'> {{=blog.blurb}} </div> {{ } }} </div> </div> </div> </div> {{ }) }} {{ if (blogPosts.length == 0) { }} <div class='s-blog-entry'> <div class='s-common-status no-posts-error'> 没有博文。 </div> </div> {{ } }} </div> {{ if (blogPosts.length > 0) { }} <div class='s-blog-col-foot'> <a class='s-blog-link s-blog-next-link' href='javascript:void(0);'> &laquo; 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Revision as of 21:21, 13 October 2016

<!DOCTYPE html>   Ellz's Site                                       
  • The Proof of Concept
    Design:
     
    Our original design includes two plasmids, one acts as a sensor module (Fig.1), another acts as an signal amplification module (Fig.2).
     
    The sensor plasmid contains pCpxP promoter which is inducible by various carbon sources including glucose, glycerol and citrate acid. Our experiment data also proved that the increase in pH could enchance the expression of pCpxP.
     
     
    Fig.1 Sensor plasmid (pLV1C3-pCpxP-LacZW-Cre)
     
     
    Fig.2 Signal amplification plasmid (pET28a-LSL-LacZW-Cre)
    Sensor plasmid uses pCpxP promoter to control the expression level of the downstream genes---genes coding for LacZ and Cre. Cre is a recombinase which recognise loxp sites and recombine the target gene based on different directions of two loxp sites which flank the target gene. When Cre recombinase has been expressed from the sensor plasmid, it will cut down two B0015 terminators on the signal amplification plasmid and recombine the plasmid (Fig.3).
     
     
    Fig.3 Plasmid recombination by Cre at loxp sites
    When the B0015 terminators downstream of T7 promoter have been cut off, genes coding for LacZ and Cre could be expressed continuously given that IPTG is in excess supply. Chain reaction will be trigger because recombined signal amplification plasmid itself could express Cre recombinase that cut off B0015 terminators on intact signal amplification plasmids. Thus, the amount of LacZ will be accumulated in an exponential rate and the expression of LacZ will be sustained even pCpxP inducers are used up.
     
    Experiment:
    Plasmid assembly method
    Genes coding for LacZ and Cre are obtained by PCR from plasmids which contain these two gene sequences with our designed primers. pCpxP promoter was synthesized by using phosphorylated single stranded oligonucleotides purchased from Invitrogen(TM). LSL(loxp-B0015-B0015-loxp) was also obtained by PCR with specially design primers. We then assembled our plasmids by using standard MCS subcloning method.
    Obstacles and changes of the original design
    Due to technical and time issues, we are not able to follow our original design. Instead, we synthesized two relatively simple plamids(Fig.4 & Fig.5) with pCpxP promoter to test for the viability of the cell-free paper-based biochip system.
     
     
    Fig.4 pLV1C3-pCpxP-GFP
     
     
    Fig.5 pLV1C3-pCpxP-LacZW
    pCpxP expression in cell
    We first transformed the plasmid pLV1C3-pCpxP-GFP into bacteria E.coli DH5α and test for the OD and RFU value under different concentrations of glucose and glycerol in order to characterize the pCpxP promoter. Based on a recent research, promoter CpxP was found to have potential in detecting a variaty of carbon sources in clinical samples[1]. However, our experimental results do not fit this research, the trend of our data generally cannot prove the inducibility of pCpxP by glucose or glycerol (Fig.6 & Fig.7).
     
     
    Fig.6 MRPU against glucose concentration (mg/dL)
     
     
    Fig.7 MRPU against glycerol concentration (mg/dL)
    MRPU (modified relative promoter activity unit) is a equation that we modified from RPU[2] in order to determine the inducibility of a particular promoter by comparing the expression unit of the promoter with the expression unit of a reference promoter --- J23119 constitutive promoter. MRPU is a relatively simple equation that use much fewer parameters than RPU and provides a much easier way to perform the calculation to give a general idea about the inducibility of a particular promoter by particular factors.
     
     
    Fig.8 Equations used to calculate MRPU Here we use pCpxP as promoter
    According to Wolfe’s study, Cpx signal transduction system, in conjunction with SigmaE and Sigma32, responds to a board spectrum of adverse environmental conditions including high pH[3]. So we then test for the expression of promoter CpxP in E.coli DH5α in a range of pH from 6 to 9. We cultured bacteria in different pH under 37℃ for 10 hours and record the RFU and OD values. Graph of MRPU against pH has been plotted (Fig.9).
     
     
    Fig.9 Graph of MRPU against pH
    The graph shows an logarithmic relationship between MRPU and pH, indicated the expression level of pCpxP increases when the pH rises. The MRPU at pH 9 is about three-folds the MRPU when pH is 6. Thus, the result shows that the expression level of pCpxP could be greatly increased when the pH has been increased in a certain range.
    pCpxP expression in S30 cell-free system
    Next, we put the pLV1C3-pCpxP-GFP and J23119-GFP plasmids into S30 cell-free system and record the RFU values of the mix after reaction for 1 hour to see if pCpxP promoter can express GFP protein in S30 cell-free system. The data proved that the pCpxP promoter can initiate the transcription in S30 cell-free system and has much higher expression level than J23119 promoter(Fig.10).
     
     
    Fig.10 Comparison between the RFU value of pCpxP-GFP and J23119-GFP plasmids in S30 cell-free system after cultured for 1 hour
    Due to time issue, we are unable to test for the expression level of pCpxP in different pH in S30 cell-free system. However, this data will be gained later and updated to our part pages.