Difference between revisions of "Team:CSU CHINA/Composite Part"

(Prototype team page)
 
Line 1: Line 1:
 
{{CSU_CHINA}}
 
{{CSU_CHINA}}
 
<html>
 
<html>
 +
<body style="position: relative" data-spy="scroll" data-offset="300" data-target="#myScrollspy">
 +
    <style>
 +
          .container{
 +
                margin-bottom:5%;
 +
                margin-top:5%;
 +
                }
 +
          .Topbox{
 +
                    width: 100%;
 +
                    min-height: 600px;
 +
                    display: -webkit-flex; /* Safari */
 +
                    display: flex;
 +
                    color:white;
 +
                    background-image: url('https://static.igem.org/mediawiki/2019/d/d3/T--CSU_CHINA--TM_BC.jpg');
 +
                    background-size: cover;
 +
                    background-repeat: no-repeat;
 +
                    background-position: center;
 +
                    justify-content: center;
 +
                    align-items: center;
 +
                }
 +
                h2{
 +
                    margin-top: 30px;
 +
                }
 +
                p{
 +
                    font-family: Times New Roman;
 +
                    text-align: justify;
 +
                    font-size:1rem!important;
 +
                }
 +
                a:hover{
 +
                    cursor: pointer;
 +
                }
 +
                .container img{
 +
                display: block;
 +
                margin: 0 auto;
 +
                box-shadow: 5px 5px 10px #888888;
 +
                }
 +
                .Teenav{
 +
                    background-color:#f3f3f3;
 +
                    border-radius: 10px;
 +
                }
 +
                .flex-column{
 +
                    position: -webkit-sticky; /* Safari */
 +
                    position: sticky;
 +
                    margin-top: 50px;
 +
                    top: 25%;
 +
                }
 +
                .Teenav li{
 +
                    margin:10px 0px;
 +
                }
 +
                .Teenav a{
 +
                    color: #197399;
 +
                    border-radius: 10px;
 +
                }
 +
                .Teenav a:hover{
 +
                    color: white;
 +
                    background-color: #2194c5;
 +
                }
 +
                #toTOP{
 +
                    position:fixed;
 +
                    right:2%;
 +
                    bottom:2%;
 +
                    border-radius:25%;
 +
                }
 +
                .ts {
 +
                    margin-bottom: 0px;
 +
                    margin-top: 5px;
 +
                    text-align: center!important;
 +
                }
 +
    </style>
 +
    <div class="container-fluid Topbox" id="Top">
 +
            <h1 class="display-3" style="color:white">Composite Part</h1>
 +
    </div>
 +
    <div class="container">
 +
        <div class="row">
 +
            <nav class="col-sm-2"id="myScrollspy">
 +
                <ul class="nav flex-column Teenav">
 +
                        <li class="nav-item">
 +
                                <a class="nav-link" href="#Overview">Overview</a>
 +
                            </li>
 +
                            <li class="nav-item">
 +
                              <a class="nav-link" href="#Module1">Module1</a>
 +
                            </li>
 +
                            <li class="nav-item">
 +
                              <a class="nav-link" href="#Module2">Module2</a>
 +
                            </li>
 +
                            <li class="nav-item">
 +
                              <a class="nav-link" href="#Module3">Module3</a>
 +
                            </li>
 +
                            <li class="nav-item">
 +
                                    <a class="nav-link" href="#References">References</a>
 +
                            </li>
 +
                </ul>
 +
            </nav>
 +
            <div class="col-sm-10">
 +
                <div id="mRNA" style="width: 100%; margin-bottom: 30px;">
 +
                    <div class="card">
 +
                            <div class="card-header">
 +
                                <a class="card-link" data-toggle="collapse" href="#mRNA1" style="color: black">Composite Part</a>
 +
                            </div>
 +
                            <div id="mRNA1" class="collapse" data-parent="#mRNA">
 +
                                <div class="card-body">
 +
                                    <figure>
 +
                                        <center>
 +
                                                <div>
 +
                                                        <table class="table table-hover table-responsive-sm">
 +
                                                                <thead>
 +
                                                                    <tr>
 +
                                                                            <td>Section</td>
 +
                                                                            <td>Name</td>
 +
                                                                            <td>Type</td>
 +
                                                                            <td>Description</td>
 +
                                                                    </tr>
 +
                                                                    </thead>
 +
                                                                    <tbody>
 +
                                                                        <tr>
 +
                                                                            <th>Module1</th>
 +
                                                                            <td><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2908671">BBa_K2908671</a></td>
 +
                                                                            <td>Plasmid</td>
 +
                                                                            <td>pLN431-s(GATA3)p-GAD-miR101-BS</td>
 +
                                                                        </tr>
 +
                                                                        <tr>
 +
                                                                            <th rowspan="3">Module2</th>
 +
                                                                            <td><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2908679">BBa_K2908679</a></td>
 +
                                                                            <td>Plasmid</td>
 +
                                                                            <td>s(ESR1)p-miR141spe-EGFP</td>
 +
                                                                        </tr>
 +
                                                                        <tr>
 +
                                                                            <td><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2908680">BBa_K2908680</a></td>
 +
                                                                            <td>Plasmid</td>
 +
                                                                            <td>s(ESR1)p-miR148bspe-EGFP </td>
 +
                                                                        </tr>
 +
                                                                        <tr>
 +
                                                                            <td><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2908681">BBa_K2908681</a></td>
 +
                                                                            <td>Plasmid</td>
 +
                                                                            <td>s(ESR1)p-miR101spe-EGFP</td>
 +
                                                                        </tr>
 +
                                                                        <tr>
 +
                                                                            <th>Module3</th>
 +
                                                                            <td><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2908670">BBa_K2908670</a></td>
 +
                                                                            <td>Plasmid</td>
 +
                                                                            <td>pLN431-G8p-yCD-miR101BS</td>
 +
                                                                        </tr>
 +
                                                                    </tbody>
 +
                                                            </table>
 +
                                                </div>
 +
                                        </center>
 +
                                    </figure>
 +
                                </div>
 +
                            </div>
 +
                        </div>
 +
                    <div id="Overview">
 +
                        <h2>Overview</h2>
 +
                        <h3>√ Golden Gate assay to construct the plasmids</h3>
 +
                        <p>First step: Construction of pocket plasmid </p>
 +
                        <p>Next: ‘Golden Gate’</p>
 +
                        <p>In 2008, Engler etc. reported a kind of cloned strategy through IIS restriction enzymes , called IIS cloning,  namely ‘Golden Gate’ cloning. Type IIS restriction enzyme can specifically identify the target sites on double-stranded DNA, and non-specifically cut the DNA double-stranded downstream of the target sites to produce a cohesive end at the 5 'or 3' end of the DNA double-stranded [1]. </p>
 +
                        <p>"Golden Gate" cloning method is in the same reaction system, using IIS type restriction endonuclease, cut beyond recognition site of DNA, DNA fragments containing cohesive end, at the same time by ligase connect several DNA fragments according to the established order, and spliced into excluding enzyme recognition site of DNA fragments, is like a linear puzzles are correctly spliced together, make the multiple objective DNA fragments in the order set to realize "seamless connection”.</p>                   
 +
                    </div>
 +
                    <div id="Module1">
 +
                        <h2 >Module1</h2>
 +
                        <img src="https://static.igem.org/mediawiki/2019/6/6a/T--CSU_CHINA--CP_Module1.png" class="mx-auto" style="width:60% ">
 +
                        <p class="ts"><b>pLN431-s(GATA3)p-GAD-miR101BD (BBa_K2908671)</b></p>
 +
                        <p>We use the pLN431[2] as the back-clone of the Module1, then we cut the vector and an insert sequence (lacZp+lacZ) with MluI and NotI to form the pocket plasmid.</p>
 +
                        <div class="row">
 +
                                <div class="col-sm-6">
 +
                                <img src="https://static.igem.org/mediawiki/2019/e/e0/T--CSU_CHINA--CP_Module1_KDXQ.png" style="width:90%;margin-top:100px;">
 +
                                <p class="ts"><b>Module1 口袋质粒构建</b></p>
 +
                                </div>
 +
                                <div class="col-sm-6">
 +
                                <img src="https://static.igem.org/mediawiki/2019/8/8b/T--CSU_CHINA--CP_Module1_KD.png" class="mx-auto" style="width:90% ">
 +
                                <p class="ts"><b>Module1 口袋质粒 详细</b></p>
 +
                                </div> 
 +
                        </div>
 +
                        <p>Supplementary figure: </p>
 +
                        <img src="https://static.igem.org/mediawiki/2019/2/26/T--CSU_CHINA--CP_lacZ.png" class="mx-auto" style="width: 60% ">
 +
                        <p>We designed the BsmBI restriction digest location on the 5’ and 3’ top of the inserted sequence, the following figure shows the pattern of construction method.</p>
 +
                        <img src="https://static.igem.org/mediawiki/2019/9/9f/T--CSU_CHINA--CP_Module1-Golden_Gate.png" class="mx-auto" style="width: 60% ">
 +
                        <p class="ts"><b>Module1-Golden Gate</b></p>
 +
                    </div>
 +
                    <div  id="Module2">
 +
                        <h2>Module2</h2>
 +
                        <img src="https://static.igem.org/mediawiki/2019/6/68/T--CSU_CHINA--CP_Module2-141spe.png" class="mx-auto" style="width: 60% ">
 +
                        <p class="ts"><b>p(ESR1)p-miR141spe-EGFP (BBa_K2908672)</b></p>
 +
                        <img src="https://static.igem.org/mediawiki/2019/9/93/T--CSU_CHINA--CP_Module2-148bspe.png" class="mx-auto" style="width: 60% ">
 +
                        <p class="ts"><b>p(ESR1)p-miR148bspe-EGFP (BBa_K2908674)</b></p>
 +
                        <img src="https://static.igem.org/mediawiki/2019/b/bb/T--CSU_CHINA--CP_Module2-101spe.png" class="mx-auto" style="width: 60% ">
 +
                        <p class="ts"><b>p(ESR1)p-miR101spe-EGFP (BBa_K2908675)</b></p>
 +
                        <p>The construction method is the same as module1, only except the paired sequence of each inserted fragment, which shows in the following picture.</p>
 +
                        <img src="https://static.igem.org/mediawiki/2019/5/5b/T--CSU_CHINA--CP_Module2-Golden_Gate.png" class="mx-auto" style="width: 60% ">
 +
                        <p class="ts"><b>Module2-Golden Gate</b></p>
 +
                    </div>
 +
                    <div id="Module3">
 +
                        <h2>Module3</h2>
 +
                        <img src="https://static.igem.org/mediawiki/2019/f/f5/T--CSU_CHINA--CP_Module3.png" class="mx-auto" style="width: 60% ">
 +
                        <p class="ts"><b>pLN431-G8p-yCD-miR101BD (BBa_K2908670)</b></p>
 +
                    </div>
 +
                    <div id="References">
 +
                            <h2>References</h2>
 +
                            <p>[1] Waclaw Szybalski”, Sun C. Kim”, Noaman Hasan” and Anna J. Podhajskab, Class IIS restriction enzymes, a review.1991, Gene, 100: 13-26.</p>
 +
                            <p>[2] Lior Nissim, Ming-Ru Wu, Erez Pery, Adina Binder-Nissim, Hiroshi I. Suzuki, Doron Stupp, Claudia Wehrspaun, Yuval Tabach, Phillip A. Sharp, and Timothy K. Lu, Cell, (2017),Synthetic RNA-Based Immunomodulatory Gene Circuits for Cancer Immunotherapy.</p>
 +
                    </div>
 +
            </div>
  
−
 
+
        </div>
−
 
+
    </div>
−
<div class="column full_size judges-will-not-evaluate">
+
    <a href="#Top" class="btn btn-info" id="toTOP" role="button">▲</a>
−
<h3>★  ALERT! </h3>
+
</body>
−
<p>This page is used by the judges to evaluate your team for the <a href="https://2019.igem.org/Judging/Medals">medal criterion</a> or <a href="https://2019.igem.org/Judging/Awards"> award listed below</a>. </p>
+
−
<p> Delete this box in order to be evaluated for this medal criterion and/or award. See more information at <a href="https://2019.igem.org/Judging/Pages_for_Awards"> Instructions for Pages for awards</a>.</p>
+
−
</div>
+
−
 
+
−
 
+
−
<div class="clear"></div>
+
−
 
+
−
 
+
−
 
+
−
 
+
−
 
+
−
<div class="column full_size">
+
−
<h1>Composite Parts</h1>
+
−
 
+
−
 
+
−
<p>
+
−
A composite part is a functional unit of DNA consisting of two or more basic parts assembled together. <a href="http://parts.igem.org/wiki/index.php/Part:BBa_I13507">BBa_I13507</a> is an example of a composite part, consisting of an RBS, a protein coding region for a red fluorescent protein, and a terminator.
+
−
</p>
+
−
 
+
−
<p>New composite BioBrick devices can be made by combining existing BioBrick Parts (like Inverters, Amplifiers, Smell Generators, Protein Balloon Generators, Senders, Receivers, Actuators, and so on).</p>
+
−
</div>
+
−
 
+
−
<div class="column full_size">
+
−
<div class="highlight decoration_background">
+
−
<h3>Note</h3>
+
−
<p>This page should list all the composite parts your team has made during your project and include direct links to your Parts main pages on the Registry. <b>You must add all characterization information for your parts on Parts Main Page on the Registry.</b> You should <b>not</b> put characterization information on this page. Remember judges will only look at the first part in the list for the Best Composite Part award, so put your best part first!</p>
+
−
</div>
+
−
</div>
+
−
 
+
−
 
+
−
 
+
−
<div class="column full_size">
+
−
<h3>Best Composite Part Special Prize</h3>
+
−
 
+
−
<p>To be eligible for this award, this part <b>must be well documented on the part's Main Page on the Registry</b>. If you have a part you wish to nominate your team for this <a href="https://2019.igem.org/Judging/Awards">special prize</a>, make sure you add your part number to your <a href="https://2019.igem.org/Judging/Judging_Form">judging form</a> and delete the alert box at the top of this page.
+
−
 
+
−
<br><br>
+
−
<b>Please note:</b> Judges will only look at the first part number you list, so please only enter ONE (1) part number in the judging form for this prize. </p>
+
−
 
+
−
</div>
+
−
 
+
 
</html>
 
</html>

Revision as of 08:20, 20 October 2019

loading……

Composite Part

Composite Part
Section Name Type Description
Module1 BBa_K2908671 Plasmid pLN431-s(GATA3)p-GAD-miR101-BS
Module2 BBa_K2908679 Plasmid s(ESR1)p-miR141spe-EGFP
BBa_K2908680 Plasmid s(ESR1)p-miR148bspe-EGFP
BBa_K2908681 Plasmid s(ESR1)p-miR101spe-EGFP
Module3 BBa_K2908670 Plasmid pLN431-G8p-yCD-miR101BS

Overview

√ Golden Gate assay to construct the plasmids

First step: Construction of pocket plasmid

Next: ‘Golden Gate’

In 2008, Engler etc. reported a kind of cloned strategy through IIS restriction enzymes , called IIS cloning, namely ‘Golden Gate’ cloning. Type IIS restriction enzyme can specifically identify the target sites on double-stranded DNA, and non-specifically cut the DNA double-stranded downstream of the target sites to produce a cohesive end at the 5 'or 3' end of the DNA double-stranded [1].

"Golden Gate" cloning method is in the same reaction system, using IIS type restriction endonuclease, cut beyond recognition site of DNA, DNA fragments containing cohesive end, at the same time by ligase connect several DNA fragments according to the established order, and spliced into excluding enzyme recognition site of DNA fragments, is like a linear puzzles are correctly spliced together, make the multiple objective DNA fragments in the order set to realize "seamless connection”.

Module1

pLN431-s(GATA3)p-GAD-miR101BD (BBa_K2908671)

We use the pLN431[2] as the back-clone of the Module1, then we cut the vector and an insert sequence (lacZp+lacZ) with MluI and NotI to form the pocket plasmid.

Module1 口袋质粒构建

Module1 口袋质粒 详细

Supplementary figure:

We designed the BsmBI restriction digest location on the 5’ and 3’ top of the inserted sequence, the following figure shows the pattern of construction method.

Module1-Golden Gate

Module2

p(ESR1)p-miR141spe-EGFP (BBa_K2908672)

p(ESR1)p-miR148bspe-EGFP (BBa_K2908674)

p(ESR1)p-miR101spe-EGFP (BBa_K2908675)

The construction method is the same as module1, only except the paired sequence of each inserted fragment, which shows in the following picture.

Module2-Golden Gate

Module3

pLN431-G8p-yCD-miR101BD (BBa_K2908670)

References

[1] Waclaw Szybalski”, Sun C. Kim”, Noaman Hasan” and Anna J. Podhajskab, Class IIS restriction enzymes, a review.1991, Gene, 100: 13-26.

[2] Lior Nissim, Ming-Ru Wu, Erez Pery, Adina Binder-Nissim, Hiroshi I. Suzuki, Doron Stupp, Claudia Wehrspaun, Yuval Tabach, Phillip A. Sharp, and Timothy K. Lu, Cell, (2017),Synthetic RNA-Based Immunomodulatory Gene Circuits for Cancer Immunotherapy.

▲