|
|
| Line 7: |
Line 7: |
| | <meta name="renderer" content="webkit"> | | <meta name="renderer" content="webkit"> |
| | <!-- 极速模式 --> | | <!-- 极速模式 --> |
| − | <title>Application</title> | + | <title>Description</title> |
| | <link rel="stylesheet" href="https://2016.igem.org/Team:HUST-China/css/bootstrap-theme?action=raw&ctype=text/css"> | | <link rel="stylesheet" href="https://2016.igem.org/Team:HUST-China/css/bootstrap-theme?action=raw&ctype=text/css"> |
| | <link rel="stylesheet" href="https://2016.igem.org/Team:HUST-China/css/bootstrap?action=raw&ctype=text/css"> | | <link rel="stylesheet" href="https://2016.igem.org/Team:HUST-China/css/bootstrap?action=raw&ctype=text/css"> |
| Line 14: |
Line 14: |
| | <script type="text/javascript" src="https://2016.igem.org/Team:HUST-China/js/bootstrap?action=raw&ctype=text/javascript"></script> | | <script type="text/javascript" src="https://2016.igem.org/Team:HUST-China/js/bootstrap?action=raw&ctype=text/javascript"></script> |
| | <script type="text/javascript" src="https://2016.igem.org/Team:HUST-China/js/index?action=raw&ctype=text/javascript"></script> | | <script type="text/javascript" src="https://2016.igem.org/Team:HUST-China/js/index?action=raw&ctype=text/javascript"></script> |
| | + | <!-- <script src="../js/jquery.tocible.min.js"></script> --> |
| | <script type="text/javascript" src="https://2016.igem.org/Team:HUST-China/js/tocible?action=raw&ctype=text/javascript"></script> | | <script type="text/javascript" src="https://2016.igem.org/Team:HUST-China/js/tocible?action=raw&ctype=text/javascript"></script> |
| − | <style>
| + | <style> |
| | html { | | html { |
| | overflow-x: hidden; | | overflow-x: hidden; |
| Line 72: |
Line 73: |
| | box-sizing: content-box; | | box-sizing: content-box; |
| | } | | } |
| | + | |
| | #sideMenu .mainMenu_toggle, | | #sideMenu .mainMenu_toggle, |
| | #sideMenu .mainMenu_Title, | | #sideMenu .mainMenu_Title, |
| | #sideMenu .sideMenuTitle, | | #sideMenu .sideMenuTitle, |
| − | #sideMenu .expand_subMenus{ | + | #sideMenu .expand_subMenus { |
| | box-sizing: content-box; | | box-sizing: content-box; |
| | } | | } |
| Line 137: |
Line 139: |
| | <div class="banner-top"> | | <div class="banner-top"> |
| | <ul class="nav banner-nav"> | | <ul class="nav banner-nav"> |
| − | <li><a class="active" href="#">HOME</a></li> | + | <li><a href="https://2016.igem.org/Team:HUST-China">HOME</a></li> |
| − | <li class="dropdown1"><a class="down-scroll" href="#">PROJECT</a> | + | <li class="dropdown1"><a class="down-scroll active" href="#">PROJECT</a> |
| | <ul class="dropdown2"> | | <ul class="dropdown2"> |
| | <li><a href="https://2016.igem.org/Team:HUST-China/project/background">Background</a></li> | | <li><a href="https://2016.igem.org/Team:HUST-China/project/background">Background</a></li> |
| Line 148: |
Line 150: |
| | </ul> | | </ul> |
| | </li> | | </li> |
| − | <li class="dropdown1"><a href="#">WETLAB</a> | + | <li class="dropdown1"><a href="#" class="down-scroll">WETLAB</a> |
| | <ul class="dropdown2"> | | <ul class="dropdown2"> |
| | <li><a href="https://2016.igem.org/Team:HUST-China/Experiments">Experiments</a></li> | | <li><a href="https://2016.igem.org/Team:HUST-China/Experiments">Experiments</a></li> |
| Line 158: |
Line 160: |
| | </li> | | </li> |
| | <li class="dropdown1"><a class="down-scroll" href="https://2016.igem.org/Team:HUST-China/Model">MODELING</a> | | <li class="dropdown1"><a class="down-scroll" href="https://2016.igem.org/Team:HUST-China/Model">MODELING</a> |
| − | <!-- <ul class="dropdown2"> | + | |
| − | <li><a href=" #"> Mag-receiver</a></li> | + | <li><a href=" https://2016.igem.org/Team:HUST-China/Model"> Overview</a></li> |
| − | <li><a href=" #"> Heating</a></li> | + | <li><a href=" https://2016.igem.org/Team:HUST-China/Model/model-pro"> Prokaryotic circuit</a></li> |
| − | <li><a href=" #"> Thermo-regulator</a></li> | + | <li><a href=" https://2016.igem.org/Team:HUST-China/Model/model-euk"> Eukaryotic circuit</a></li> |
| − | </ul> -->
| + | <li><a href="https://2016.igem.org/Team:HUST-China/Model/model-app">Application circuit</a></li> |
| − | </li>
| + | </ul> |
| | + | </li> |
| | <li class="dropdown1"><a class="down-scroll" href="">PARTS</a> | | <li class="dropdown1"><a class="down-scroll" href="">PARTS</a> |
| | <ul class="dropdown2"> | | <ul class="dropdown2"> |
| Line 205: |
Line 208: |
| | <div class="ref"></div> | | <div class="ref"></div> |
| | <article> | | <article> |
| | + | <p>This is a stage where Syn-bio can make a difference. Our team was inspired by the undergraduate finalist ”Noise” and their excellent work for synthetic biology. And this year, we wanted to participate and contribute too.</p> |
| | <!-- h2一级标题 --> | | <!-- h2一级标题 --> |
| − | <h2>OVERVIEW</h2> | + | <h2>Lactose intolerance</h2> |
| | <!-- 文字 --> | | <!-- 文字 --> |
| | <p> | | <p> |
| − | This year, we want to offer a handy, adjustable and useful toolkit to everyone working on Synthetic Biology--Signal Filter. The core circuit is based on a positive feedback bi-stable or tri-stable system with the capacity of reducing noise and converting pulse signal into robust and persistent signal. To adapt to different experimental requirements, we also developed two versions of our Signal Filter--prokaryotic one and eukaryotic one. | + | <strong>At the beginning</strong>, we learned about previous projects from the websites, and we realized that, when dealing with real-world problems, issues like gene regulation, expression efficiency and system robustness all matter. When creating a great functional prototype, one should pay attention to all the details. And some of the new comers to synthetic biology may find it difficult to deal with. So we put it in our way: why not provide some gene expression kits to iGEMers so that they no longer need to worry about building circuit, but focus on the key problem. |
| | </p> | | </p> |
| | + | <p> |
| | + | We came up with different versions of gene expression circuit: bacteriophage lambda, kinases reaction pathway, ribo-switch, RNAi and so on. We made efforts on the circuit construction and valid characterization data while users mainly focus on the input and output which are highly concerned with real-world problems. In this manner, our kit can serve as a useful tool to save their time and energy. On the whole, our theme is to offer bricks to help others build their own project. |
| | + | </p> |
| | + | <p>And this summer we <strong>stepped a little forward</strong> to our goals. We worked out two versions of gene expression switch----a prokaryote tri-stable version derived from bacteriophage lambda and eukaryote bi-stable version based on ABA-response pathway. |
| | + | </p> |
| | + | <p> |
| | + | <strong>In the tri-stable gene expression switch,</strong> users should define two input and related output gene signals. The logic gate below can illustrate the circuit better: |
| | + | </p> |
| | + | <img src="https://static.igem.org/mediawiki/2016/7/70/T--HUST-China--Logic-gate.jpg" alt=""> |
| | + | <p>The circuit can rapidly reach its stable state because of positive feedback. And users can adjust its threshold too. </p> |
| | <!-- h3二级标题 --> | | <!-- h3二级标题 --> |
| | <!-- <h3>SubHeading</h3> | | <!-- <h3>SubHeading</h3> |
| Line 216: |
Line 230: |
| | <h3>Hello world</h3> | | <h3>Hello world</h3> |
| | <p>fdwfewf</p> --> | | <p>fdwfewf</p> --> |
| − | <h2>Prokaryotic version:</h2> | + | <p>And <strong>for more application examples:</strong></p> |
| − | <p> | + | <p>We provide a solution to lactose intolerance based on this toolkit.</p> |
| − | It is a tri-stable system adapted from bacteriophage λ operon. In the operon, promoter RE is activated by the transcriptional activator CII. Ftsh is an ATP-dependent host metalloprotease which will normally degrade CII, while CIII serves as an inhibitor of Ftsh to free CII. And CI can function as an inhibitor to block pR, while Cro can bind pRE to stop the downstream gene’s expression. The positive feedback control under pRE is used to enhance pulse signal 1 and convert it into a robust and stable signal.
| + | <p>The input of signals are achieved by two promoters: plac(lactose inducible) and patp2(base inducible). As for the output,we set the gene of interest 1 as iLDH and gene of interest 2 as beta-galactosidase.iLDH can transform lactic acid into pyruvate whileβ-galactosidase can degrade lactose. In this way, the system can provide a promising way for the treatment of lactose intorlance.</p> |
| − | </p>
| + | <img src="https://static.igem.org/mediawiki/2016/c/c8/T--HUST-China--description-app.png" alt=""> |
| − | <!-- 图片 -->
| + | <p><strong>In our bi-stable switch</strong>, users can define an open signal and off signal to control the expression of target gene . The most attractive feature of the circuit is switch efficiency, the mechanism of which is cascade reaction.</p> |
| − | <img src="https://static.igem.org/mediawiki/2016/8/88/T--HUST-China--Description-Fig-Eukaryote.png" alt="" class="img-responsive"> | + | <img src="https://static.igem.org/mediawiki/2016/8/83/T--HUST-China--description-logic.png" alt=""> |
| − | <p class="text-center">Fig*: Circuit of prokaryotic version serving as a tri-stable signal filter</p> | + | |
| − | <!-- <h2>Creation based on Signal Filter</h2> -->
| + | |
| − | <!-- 文字 -->
| + | |
| − | <p>
| + | |
| − | When signal 1 comes, pRE will be activated, expressing Cro and CII. At the same time, the constitutively produced CIII will guarantee enough CII to enhance the transcription downstream the pRE. Thus,there forms a positive feedback loop to direct a fast and strong expression of Cro. A quantity of Cro can repress the transcription under pRM by binding to Cro binding site and blocks gene of interest 2’s expression, so it turns to a stable state of expressing gene of interest 1.
| + | |
| − | </p>
| + | |
| − | <!-- <img src="" alt=""> -->
| + | |
| − | <p>When signal 2 comes, under a certain inducible promoter, CI will be expressed and then binds to CI binding site, blocking the expression of gene of interest 1 and cIII. With CIII’s reduction, FtsH gradually degrades CII and interrupts the stable state. Therefore, Cro’s expression will immediately drop down, allowing the expression of gene of interest 2. So the system turns to another stable state.
| + | |
| − | </p>
| + | |
| − | <p>
| + | |
| − | If there is no input signal, both genes of interest will be expressed. And if both of the two signals exist, the expression state will depend on the intensity of the initial input signal.
| + | |
| − | </p>
| + | |
| − | <h2>Eukaryotic version:</h2>
| + | |
| − | <p>It is a bi-stable system derived from Arabidopsis thaliana stress response system. Enzyme catalysis is the core of this design to increase the efficiency of state transition.
| + | |
| − | </p>
| + | |
| − | <img src="https://static.igem.org/mediawiki/2016/1/1e/T--HUST-China--Description-Fig-prokaryote.png" alt="" class="img-responsive"> | + | |
| − | <p class="text-center">Fig*: Circuit of Eukaryotic version serving as a bi-stable signal filter</p>
| + | |
| − | <p>Clade A protein phosphatases type 2C(PP2CA) and SUCROSE NONFERMENTING1-RELATED SUBFAMILY2(SnRK2s) protein kinase are both components of Abscisic acid signaling network in Arabidopsis thaliana. ABF2 is a leucine zipper transcription factor which basically binds to ABA-response element (ABRE). ABF2 can be phosphorylated by SnRK2s and efficiently dephosphorylated by PP2CA. Also, in the absence of Abscisic acid, SnRK2 kinases will be inactivated by PP2Cs and thus, efficiently shut down the system.
| + | |
| − | </p>
| + | |
| − | <p>When signal 1(ON) comes, promoter RD29A drives the expression of SnRK2.2, which can phosphorylate ABF2 (constitutively express). Comparing to protein co-facter association, enzyme catalysis can produce a large quantity of phosphorylated ABF2 in a short time, then enhances pRD29A to turn on the gene of interest’s expression. When signal 2(OFF) comes, gene PP2CA expresses, dephosphorylating ABF2 and inactivating SnRK2.2, thus turning the system into OFF state.
| + | |
| − | </p>
| + | |
| | </article> | | </article> |
| | </div> | | </div> |
