Team/CINVESTAV-IPN-UNAM MX/Construction.htm

From 2012.igem.org

(Difference between revisions)
(Created page with " <html xmlns="http://www.w3.org/1999/xhtml"><head> <title>Home</title> <link rel="stylesheet" href="https://dl.dropbox.com/u/199422/Cinvestav/style.css" /> <meta http-equiv="C...")
 
(21 intermediate revisions not shown)
Line 1: Line 1:
 +
<html xmlns="http://www.w3.org/1999/xhtml">
 +
<head>
 +
<meta http-equiv="content-type" content="text/html; charset=utf-8" />
 +
<title>Rho</title>
 +
<style type="text/css">
-
<html xmlns="http://www.w3.org/1999/xhtml"><head>
+
#top-section {
-
<title>Home</title>
+
position: relative;
 +
height: 20px;
 +
width: 975px;
 +
margin: 0 auto 0 auto;
 +
border-left: hidden;
 +
border-right: hidden;
 +
border-bottom: hidden;
 +
}
-
<link rel="stylesheet" href="https://dl.dropbox.com/u/199422/Cinvestav/style.css" />
+
* {
 +
        margin: 0;
 +
        padding: 0;
 +
}
 +
body {
 +
        background: #FFFFFF url(https://static.igem.org/mediawiki/2012/f/f3/Fondo_bod.jpg) repeat-x;
 +
        text-align: justify;
 +
        font-family: Georgia, "Times New Roman", Times, serif;
 +
        font-size: 12px;
 +
        color: #000000;
 +
}
-
<meta http-equiv="Content-Type" content="text/html; charset=utf-8" />
+
h1, h2, h3 {
-
<style type="text/css">td img {display: block;}</style>
+
        color: #333333;
 +
}
-
<!--Fireworks CS6 Dreamweaver CS6 target.  Created Tue Sep 25 17:05:18 GMT-0500 (Central Daylight Time (Mexico)) 2012-->
+
h1, h2 {
-
</head>
+
        margin-bottom: 20px;
-
<body bgcolor="#ffffff">
+
        font-weight: normal;
-
<table style="display: inline-table;" border="0" cellpadding="0" cellspacing="0" width="950" bgcolor="">
+
}
-
<!-- fwtable fwsrc="INDEX.fw.png" fwpage="Page 1" fwbase="principal.jpg" fwstyle="Dreamweaver" fwdocid = "927795023" fwnested="0" -->
+
-
  <tr>
+
-
  <td><img src="images/spacer.gif" width="240" height="1" alt="" /></td>
+
-
  <td><img src="images/spacer.gif" width="710" height="1" alt="" /></td>
+
-
  <td><img src="images/spacer.gif" width="1" height="1" alt="" /></td>
+
-
  </tr>
+
-
<tr>
+
h1 {
-
  <td><img src="https://static.igem.org/mediawiki/2012/6/68/LOGOPRIN.jpg" width="240" height="178" alt="Logo Principal" /></td>
+
        font-size: 197%;
-
  <td bgcolor="01A79D"><ul class="nav">
+
}
-
<li>
+
 
-
<a href="/Team/CINVESTAV-IPN-UNAM_MX/home.htm">Home<span class="flecha">&#9660;</span></a>
+
h2 {
-
</li>
+
        font-size: 152%;
-
<li>
+
}
-
<a href="#">Rhodofactory<span class="flecha">&#9660;</span></a>
+
 
-
<ul>
+
h3 {
-
<li><a href="/Team/CINVESTAV-IPN-UNAM_MX/Overview.htm">Overview<span class="flecha">&#9660;</span></a></li>
+
        font-size: 100%;
-
           
+
}
-
            <li><a href="/Team/CINVESTAV-IPN-UNAM_MX/Light Response.htm">Light Response<span class="flecha">&#9660;</span></a></li>
+
 
-
            <li><a href="/Team/CINVESTAV-IPN-UNAM_MX/Oxigen Response.htm">Oxigen Response<span class="flecha">&#9660;</span></a></li>
+
p, ul, ol {
-
            <li><a href="/Team/CINVESTAV-IPN-UNAM_MX/Chassis.htm">Chassis<span class="flecha">&#9660;</span></a></li>
+
        margin-bottom: 20px;
-
            <li><a href="/Team/CINVESTAV-IPN-UNAM_MX/Perspective.htm">Perspective<span class="flecha">&#9660;</span></a></li>
+
        line-height: 1.8em;
-
+
}
-
</ul>
+
 
-
</li>
+
ul, ol {
-
<li>
+
        margin-left: 3em;
-
<a href="">Results<span class="flecha">&#9660;</span></a>
+
}
-
<ul>
+
#text2
-
<li><a href="/Team/CINVESTAV-IPN-UNAM_MX/Biobricks.htm">Biobricks<span class="flecha">&#9660;</span></a></li>
+
{
-
           
+
color: #333333;
-
            <li><a href="/Team/CINVESTAV-IPN-UNAM_MX/Notebook.htm">Notebook<span class="flecha">&#9660;</span></a></li>
+
font-size: 152%;
-
            <li><a href="/Team/CINVESTAV-IPN-UNAM_MX/Protocol.htm">Protocol<span class="flecha">&#9660;</span></a></li>
+
margin-bottom: 20px;
-
            <li><a href="/Team/CINVESTAV-IPN-UNAM_MX/palustris response.htm">R. palustris response<span class="flecha">&#9660;</span></a></li>
+
font-weight: normal;
-
            <li><a href="/Team/CINVESTAV-IPN-UNAM_MX/sphaeroides response.htm">R. sphaeroides response<span class="flecha">&#9660;</span></a></li>
+
}
-
</ul>
+
blockquote {
-
</li>
+
        padding-left: 3em;
-
+
        background: url(https://static.igem.org/mediawiki/2012/1/11/Img08.gif) no-repeat;
-
<li>
+
        font-style: italic;
-
<a href="#">IGEM Contribution<span class="flecha">&#9660;</span></a>
+
}
-
<ul>
+
 
-
<li><a href="/Team/CINVESTAV-IPN-UNAM_MX/Construction.htm">Construction<span class="flecha">&#9660;</span></a></li>
+
a:link {
-
           
+
        color: #1953A7;
-
            <li><a href="/Team/CINVESTAV-IPN-UNAM_MX/Metods.htm">Metods<span class="flecha">&#9660;</span></a></li>
+
}
-
            <li><a href="/Team/CINVESTAV-IPN-UNAM_MX/Results.htm">Results<span class="flecha">&#9660;</span></a></li>
+
 
-
+
a:active {
-
</ul>
+
        color: #ED5C0C;
-
</li>
+
}
-
<li>
+
 
-
<a href="/Team/CINVESTAV-IPN-UNAM_MX/Modelling.htm">Modelling<span class="flecha">&#9660;</span></a>
+
a:visited {
-
+
        color: #666666;
-
</li>  
+
}
-
   
+
 
-
    <li>
+
a:hover {
-
<a href="#">Team<span class="flecha">&#9660;</span></a>
+
        text-decoration: none;
-
<ul>
+
        color: #067600;
-
<li><a href="/Team/CINVESTAV-IPN-UNAM_MX/Members.htm">Members<span class="flecha">&#9660;</span></a></li>
+
}
-
           
+
 
-
            <li><a href="/Team/CINVESTAV-IPN-UNAM_MX/Institute.htm">Institute<span class="flecha">&#9660;</span></a></li>
+
img {
-
            <li><a href="/Team/CINVESTAV-IPN-UNAM_MX/Labs.htm">Labs<span class="flecha">&#9660;</span></a></li>
+
        border: none;
-
            <li><a href="/Team/CINVESTAV-IPN-UNAM_MX/Sponsor.htm">Sponsor<span class="flecha">&#9660;</span></a></li>
+
}
-
            <li><a href="/Team/CINVESTAV-IPN-UNAM_MX/Gallery.htm">Gallery<span class="flecha">&#9660;</span></a></li>
+
 
-
            <li><a href="/Team/CINVESTAV-IPN-UNAM_MX/Acknowledgments.htm">Acknowledgments<span class="flecha">&#9660;</span></a></li>
+
img.left {
-
+
        float: left;
-
</ul>
+
        margin: 0 20px 10px 0;
-
</li>
+
}
-
     <li>
+
 
-
<a href="#">Outreach<span class="flecha">&#9660;</span></a>
+
/* Menu */
-
<ul>
+
 
-
            <li><a href="/Team/CINVESTAV-IPN-UNAM_MX/Human Practice.htm">Human Practice<span class="flecha">&#9660;</span></a></li>
+
#menu {
-
            <li><a href="/Team/CINVESTAV-IPN-UNAM_MX/Safety.htm">Safety<span class="flecha">&#9660;</span></a></li>
+
        width: 770px;
-
</ul>
+
        height: 65px;
-
</li>
+
        margin: 0 auto;
-
      
+
        border-right: 1px solid #EDEBD5;
-
      
+
}
-
</ul></td>
+
 
-
  <td><img src="images/spacer.gif" width="1" height="150" alt="" /></td>
+
#menu ul {
-
  </tr>
+
        margin: 0;
-
  <tr>
+
        padding: 0;
-
  <td colspan="2" >
+
        list-style: none;
-
 
+
}
-
  <div id="main">
+
 
-
<div id="bodys_r1_c2">
+
#menu li {
 +
        display: inline;
 +
        font-size: 12px;
 +
        color:#4E4E4E;
 +
}
 +
 
 +
#menu a {
 +
        display: block;
 +
        float: left;
 +
        width: 95px;
 +
        height: 45px;
 +
        padding: 10px 0 0 0;
 +
        border-left: 1px solid #EDEBD5;
 +
        text-transform: lowercase;
 +
        text-decoration: none;
 +
        text-align: center;
 +
        font-size: 135%;
 +
        color: #BABABA;
 +
}
 +
 
 +
#menu a:hover {
 +
        background: #F7F7F4;
 +
}
 +
 
 +
#menu .active a {
 +
        background: #5dbb10;
 +
        color: #FFFFFF;
 +
}
 +
 
 +
/* Header */
 +
 
 +
#header {
 +
        width: 770px;
 +
        height: 170px;
 +
        margin: 0 auto;
 +
}
 +
 
 +
#header h1 {
 +
        float: left;
 +
        width: 220px;
 +
        height: 100px;
 +
        margin: 0;
 +
        padding: 70px 0 0 0;
 +
        background: url(https://static.igem.org/mediawiki/2012/0/0a/Img02.jpg) no-repeat;
 +
        text-align: center;
 +
        font-size: 36px;
 +
}
 +
 
 +
#header h2 {
 +
        float: right;
 +
        width: 530px;
 +
        height: 86px;
 +
        margin: 0;
 +
        background: url(https://static.igem.org/mediawiki/2012/4/4a/Img03.jpg) no-repeat;
 +
        padding: 84px 20px 0 0;
 +
        text-align: right;
 +
}
 +
 
 +
#header a {
 +
        text-transform: lowercase;
 +
        text-decoration: none;
 +
        font-style: italic;
 +
        font-weight: normal;
 +
        color: #FFFFFF;
 +
}
 +
 
 +
/* Page */
 +
 
 +
#page {
 +
        width: 770px;
 +
        margin: 0 auto;
 +
        padding: 20px 0;
 +
}
 +
#context{
 +
        float: left;
 +
        width: 570px;
 +
        padding-top:5px;
 +
        }       
 +
/* menudecontenido */
 +
 
 +
#sidebar {
 +
        float: right;
 +
        width: 160px;
 +
        padding: 15px;
 +
        background: #EDEBD5;
 +
}
 +
 
 +
#sidebar h2 {
 +
        margin: 0;
 +
        padding: 5px 15px;
 +
        color: #FFFFFF;
 +
}
 +
 
 +
#sidebar h3 {
 +
        font-size: 70%;
 +
        color: #FFFFFF;
 +
}
 +
 
 +
#sidebar ul {
 +
        margin: 0;
 +
        padding: 15px;
 +
        list-style: none;
 +
}
 +
 
 +
#sidebar li {
 +
              font-size: 14px;
 +
}
 +
 
 +
#sidebar a {
 +
        color: #FFFFFF;
 +
}
 +
 
 +
.orangebox {
 +
        background: url(https://static.igem.org/mediawiki/2012/b/b0/Img05.gif) repeat-y;
 +
}
 +
 
 +
.orangebox h2 {
 +
        background: url(https://static.igem.org/mediawiki/2012/c/c2/Img06.gif) repeat-y;
 +
}
 +
 
 +
/* Footer */
 +
 
 +
#piedepagina {
 +
        clear: both;
 +
        height: 150px;
 +
        padding: 25px 0;
 +
        background: #EDEBD5 url(https://static.igem.org/mediawiki/2012/a/a5/Img04.gif) repeat-x;
 +
}
 +
 
 +
#sponsors{
 +
 +
        float: left;
 +
        width: 85px;
 +
        padding-top:5px;
 +
        background: #EDEBD5;
 +
 +
      }
 +
 
 +
.firstHeading {
 +
margin-bottom: .1em;
 +
visibility:hidden;
 +
}
 +
 
 +
img[src*="/wiki/skins/common/images/wiki.png"]{
 +
    width:0;
 +
    height:0px;
 +
    padding-right:975px;
 +
    background:transparent url(https://2011.igem.org/wiki/skins/common/images/wiki.png) top left no-repeat;
 +
}
 +
 
 +
</style>
 +
 
 +
<meta name="keywords" content="" />
 +
<meta name="description" content="" />
 +
</head>
 +
<body>
 +
<div id="menu">
 +
  <ul>
 +
    <li><a href="home.htm">Home</a></li>
 +
    <li><a href="rhodofactory.htm">Rhodofactory</a></li>
 +
    <li class="active"><a href="promoterselection.htm">Promoter selection </a></li>
 +
    <li><a href="results.htm">results </a></li>
 +
    <li><a href="perspectives.htm">Perspectives</a></li>
 +
    <li><a href="Modelling.htm">Modeling</a></li>
 +
    <li><a href="aboutus.htm">About us</a> </li>
 +
    <li><a href="outreach.htm">outreach</a></li>
 +
  </ul>
 +
</div>
 +
<!-- end #menu -->
 +
<div id="header">
 +
  <h1><a href="#"></a></h1>
 +
  <h2><span></span></h2>
 +
</div>
 +
<!-- end #header -->
 +
<div id="page">
 +
  <div id="context">
 +
    <h1><em>Construction!</em></h1>
 +
    <p id="text2"> iGEM PROMOTER CHARACTERIZATION</p>
 +
 +
    <p>We need to use a medium strength promoter for our constructions, so we characterize different promoters  constitutive belonging to the isolated family from Anderson collection (J23101, J23102, J23104, J23107, J23108, J2311, and J23115) these promoters were assemble with GFP to determine promoter strength, using Victor X3 Multilabel Plate Reader.<br>
 +
      <br>
 +
      <img src="https://static.igem.org/mediawiki/2012/e/e8/Pfigura1.jpg" alt="figura1" width="570" height="180"></p>
 +
    <div align="center"><img src="https://static.igem.org/mediawiki/2012/0/03/Pfigura2.jpg" alt="figura2" width="394" height="179"><br>
 +
      <br>
 +
  <img src="https://static.igem.org/mediawiki/2012/3/39/PFigura3.jpg" alt="Figura3" width="566" height="328">   </div>
 +
    <br>
 +
    <div align="center"></div>
 +
    <br>
 +
    <p id="text2"> Result</p>
 +
    <p>The characterization of the BioBricks shown in the following table:</p>
 +
     <div align="center"><img src="https://static.igem.org/mediawiki/2012/0/02/PFigura4.jpg" alt="Figura4" width="349" height="157"></div>
 +
    <br>
 +
    <img src="https://static.igem.org/mediawiki/2012/5/5b/PFigura5.jpg" alt="Figura5" width="282" height="477" class="left">
 +
    <p>&nbsp;</p>
 +
    <p>&nbsp;</p>
 +
    <p>&nbsp;</p>
 +
    <p>&nbsp;</p>
 +
    <p>&nbsp;</p>
 +
    <p>The constructions that were made, were confirmed by double digestions. The following image shows a gel double digestion with XbaI and SpeI for confirmatory promoter constructs:</p>
 +
    <br>
 +
     <br>
 +
     <br>
 +
    <br>
 +
<p>The constructions were cultivated in LB petri dishes with chloramphenicol and they were observated at UV light after it’s incubation.</p>
 +
<div align="center">
 +
  <p><img src="https://static.igem.org/mediawiki/2012/7/7a/PFigura6.jpg" alt="Figura6" width="263" height="247" class="left"><img src="https://static.igem.org/mediawiki/2012/e/e4/PFigura7.jpg" alt="Figura7" width="249" height="248" > <img src="https://static.igem.org/mediawiki/2012/0/0b/PFigura8.jpg" width="397" height="384"></p><br>
</div>
</div>
-
<div id="bodys_r1_c3">
+
<p>In the following graphs there is shown the GFP expression in function of th time and the realtive promoter force. </p><img src="https://static.igem.org/mediawiki/2012/7/72/PFigura9.jpg" alt="Figura9" width="563" height="345">
-
   
+
 
-
      <img src="https://static.igem.org/mediawiki/2012/b/b6/Construction.fw.png" width="500" height="92" /> </div>
+
<p><br>
-
<div class="clearFloat"></div>
+
  With the previous results of the characterization of the promoters there is concluded that the promoter J23107, is the strongest because it produces more RPUs.<br><br>
-
<div id="bodys_r2_c2"></div>
+
After characterization, J23104 promoter was chosen for construction of the system of regulation of light and oxygen dependent.</p>
-
<div id="bodys_r2_c3">
+
<div align="center"><img src="https://static.igem.org/mediawiki/2012/0/09/PFigura10.jpg" alt="Figura10" width="567" height="419"> </div>
-
    <div class="Text">
+
  </div>
-
   
+
  <br>
-
      <p><strong>CHARACTERIZATION OF IGEM BIOPARTS DISTRIBUTION</strong></p>
+
  <div id="sidebar">
-
      <p>As part of the points required to perform well in  2012 iGEM competition we are asked to characterize some existing biopartes on  the record, so we decided to make the characterization of constitutive promoters belonging to the family isolated from a small combinatorial library (J23101 , J23102, J23104, J23107, J23108, J2311, and J23115) which were attached to GFP  to determine promoter activity, using the equipment Victor X3 Multilabel Plate Reader.</p>
+
    <div id="updates" class="orangebox">
-
<p><img src="images/Co1.jpg" width="640" height="186" /></p>
+
      <h2>Promoter Selection </h2>
-
<p>Fig 1. General scheme of one construction of the promoter expressing GFP</p>
+
      <ul>
-
<p><img src="images/cota.jpg" width="511" height="271" /></p>
+
        <li><a href="Construction.htm" target="_parent">Construction</a></li>
-
<p><img src="images/Co2.jpg" width="640" height="470" /></p>
+
        <li><a href="Metods.htm">Methods</a></li>
-
<p>Fig 2. Final construction</p>
+
      </ul>
-
<p>&nbsp;</p>
+
     </div>
     </div>
 +
  </div>
 +
  <div style="clear: both;">&nbsp;</div>
 +
</div>
 +
<!-- end #page -->
 +
<div id="piedepagina">
 +
  <p align="center"> <strong>Rhodofactory 2012</strong> </p>
 +
<div id="sponsors">
 +
  <div align="center"><img src="https://static.igem.org/mediawiki/2012/8/8a/Icytdf.png" alt="icytdf" width="90" height="82" /></div>
 +
</div>
 +
<div id="sponsors">
 +
  <div align="center"><img src="https://static.igem.org/mediawiki/2012/6/67/Osli.png" alt="osli" width="88" height="46" /></div>
 +
</div>
 +
 +
<div id="sponsors">
 +
  <div align="center"><img src="https://static.igem.org/mediawiki/2012/f/fb/Bio.png" alt="bio" width="80" height="97" /></div>
 +
</div>
 +
<div id="sponsors"><img src="https://static.igem.org/mediawiki/2012/d/d6/Fermentas.png" alt="fermentAS" width="82" height="45" /></div>
 +
<div id="sponsors">
 +
  <div align="center"><img src="https://static.igem.org/mediawiki/2012/a/a3/Cinestav.png" alt="cinestav" width="74" height="91" /></div>
 +
</div>
 +
<div id="sponsors"><img src="https://static.igem.org/mediawiki/2012/9/9b/Genscript.png" alt="genscript" width="83" height="45" /></div>
 +
<div id="sponsors">
 +
  <div align="center"><img src="https://static.igem.org/mediawiki/2012/1/16/Unam.png" alt="unam" width="83" height="93" /></div>
 +
</div>
 +
<div id="sponsors"><img src="https://static.igem.org/mediawiki/2012/c/c9/Gto.png" alt="gto" width="84" height="46" /></div>
 +
    <div id="sponsors">
 +
      <div align="center"><img src="https://static.igem.org/mediawiki/2012/f/fa/Qimica.png" alt="quimica" width="80" height="72" /></div>
     </div>
     </div>
-
<div class="clearFloat"></div>
+
<div id="sponsors"><img src="https://static.igem.org/mediawiki/2012/7/73/Valaner.png" alt="valaner" width="84" height="48" /></div>
 +
<div id="sponsors">
 +
  <div align="center"><img src="https://static.igem.org/mediawiki/2012/7/79/Ipn.png" alt="ipn" width="70" height="96" /></div>
 +
</div>
</div>
</div>
-
 
+
<!-- end #footer -->
-
    </td>
+
-
  <td><img src="images/spacer.gif" width="1" height="500" alt="" /></td>
+
-
  </tr>
+
-
</table>
+
</body>
</body>
</html>
</html>

Latest revision as of 03:34, 27 October 2012

Rho

Construction!

iGEM PROMOTER CHARACTERIZATION

We need to use a medium strength promoter for our constructions, so we characterize different promoters constitutive belonging to the isolated family from Anderson collection (J23101, J23102, J23104, J23107, J23108, J2311, and J23115) these promoters were assemble with GFP to determine promoter strength, using Victor X3 Multilabel Plate Reader.

figura1

figura2

Figura3


Result

The characterization of the BioBricks shown in the following table:

Figura4

Figura5

 

 

 

 

 

The constructions that were made, were confirmed by double digestions. The following image shows a gel double digestion with XbaI and SpeI for confirmatory promoter constructs:





The constructions were cultivated in LB petri dishes with chloramphenicol and they were observated at UV light after it’s incubation.

Figura6Figura7


In the following graphs there is shown the GFP expression in function of th time and the realtive promoter force.

Figura9


With the previous results of the characterization of the promoters there is concluded that the promoter J23107, is the strongest because it produces more RPUs.

After characterization, J23104 promoter was chosen for construction of the system of regulation of light and oxygen dependent.

Figura10

 

Rhodofactory 2012

icytdf
osli
bio
fermentAS
cinestav
genscript
unam
gto
quimica
valaner
ipn

Retrieved from "http://2012.igem.org/Team/CINVESTAV-IPN-UNAM_MX/Construction.htm"