Team:NTNU Trondheim/Project

From 2014.igem.org

(Difference between revisions)
 
(28 intermediate revisions not shown)
Line 3: Line 3:
<meta http-equiv="Content-Type" content="text/html; charset=utf-8" />
<meta http-equiv="Content-Type" content="text/html; charset=utf-8" />
<meta name="generator" content="MediaWiki 1.16.5" />
<meta name="generator" content="MediaWiki 1.16.5" />
-
<link rel="alternate" type="application/x-wiki" title="Edit" href="/wiki/index.php?title=WikitemplateB_project&amp;action=edit" />
 
-
<link rel="edit" title="Edit" href="/wiki/index.php?title=WikitemplateB_project&amp;action=edit" />
 
<link rel="shortcut icon" href="/favicon.ico" />
<link rel="shortcut icon" href="/favicon.ico" />
-
<link rel="search" type="application/opensearchdescription+xml" href="/wiki/opensearch_desc.php" title="2014.igem.org (en)" />
+
<link rel="search" type="application/opensearchdescription+xml" href="/wiki/opensearch_desc.php" title="2013.igem.org (en)" />
-
<link title="Creative Commons" type="application/rdf+xml" href="/wiki/index.php?title=WikitemplateB_project&amp;action=creativecommons" rel="meta" />
+
<link title="Creative Commons" type="application/rdf+xml" href="/wiki/index.php?title=Team:NTNU_Trondheim/Team&amp;action=creativecommons" rel="meta" />
-
<link rel="copyright" href= "http://creativecommons.org/licenses/by/3.0/" />
+
<link rel="copyright" href="http://creativecommons.org/licenses/by/3.0/" />
-
<link rel="alternate" type="application/atom+xml" title="2014.igem.org Atom feed" href="/wiki/index.php?title=Special:RecentChanges&amp;feed=atom" /> <title>WikitemplateB project - 2014.igem.org</title>
+
<link rel="alternate" type="application/atom+xml" title="2013.igem.org Atom feed" href="/wiki/index.php?title=Special:RecentChanges&amp;feed=atom" /> <title>Team:NTNU_Trondheim/notebook - 2014.igem.org</title>
<style type="text/css" media="screen, projection">/*<![CDATA[*/
<style type="text/css" media="screen, projection">/*<![CDATA[*/
@import "/wiki/skins/common/shared.css?270";
@import "/wiki/skins/common/shared.css?270";
Line 32: Line 30:
wgVariantArticlePath=false,
wgVariantArticlePath=false,
wgActionPaths={},
wgActionPaths={},
-
wgServer="https://2014.igem.org",
+
wgServer="https://2013.igem.org",
wgCanonicalNamespace="",
wgCanonicalNamespace="",
wgCanonicalSpecialPageName=false,
wgCanonicalSpecialPageName=false,
wgNamespaceNumber=0,
wgNamespaceNumber=0,
-
wgPageName="WikitemplateB_project",
+
wgPageName="Team:Cornell/notebook",
-
wgTitle="WikitemplateB project",
+
wgTitle="Team:Cornell/notebook",
wgAction="view",
wgAction="view",
-
wgArticleId=1048,
+
wgArticleId=3451,
wgIsArticle=true,
wgIsArticle=true,
-
wgUserName="EivindD",
+
wgUserName=null,
-
wgUserGroups=["*", "user", "autoconfirmed"],
+
wgUserGroups=null,
wgUserLanguage="en",
wgUserLanguage="en",
wgContentLanguage="en",
wgContentLanguage="en",
wgBreakFrames=false,
wgBreakFrames=false,
-
wgCurRevisionId=4634,
+
wgCurRevisionId=360799,
wgVersion="1.16.5",
wgVersion="1.16.5",
wgEnableAPI=true,
wgEnableAPI=true,
Line 53: Line 51:
wgDigitTransformTable=["", ""],
wgDigitTransformTable=["", ""],
wgMainPageTitle="Main Page",
wgMainPageTitle="Main Page",
-
wgFormattedNamespaces={"-2": "Media", "-1": "Special", "0": "", "1": "Talk", "2": "User", "3": "User talk", "4": "2014.igem.org", "5": "2014.igem.org talk", "6": "File", "7": "File talk", "8": "MediaWiki", "9": "MediaWiki talk", "10": "Template", "11": "Template talk", "12": "Help", "13": "Help talk", "14": "Category", "15": "Category talk"},
+
wgFormattedNamespaces={"-2": "Media", "-1": "Special", "0": "", "1": "Talk", "2": "User", "3": "User talk", "4": "2013.igem.org", "5": "2013.igem.org talk", "6": "File", "7": "File talk", "8": "MediaWiki", "9": "MediaWiki talk", "10": "Template", "11": "Template talk", "12": "Help", "13": "Help talk", "14": "Category", "15": "Category talk"},
-
wgNamespaceIds={"media": -2, "special": -1, "": 0, "talk": 1, "user": 2, "user_talk": 3, "2014.igem.org": 4, "2014.igem.org_talk": 5, "file": 6, "file_talk": 7, "mediawiki": 8, "mediawiki_talk": 9, "template": 10, "template_talk": 11, "help": 12, "help_talk": 13, "category": 14, "category_talk": 15, "image": 6, "image_talk": 7},
+
wgNamespaceIds={"media": -2, "special": -1, "": 0, "talk": 1, "user": 2, "user_talk": 3, "2013.igem.org": 4, "2013.igem.org_talk": 5, "file": 6, "file_talk": 7, "mediawiki": 8, "mediawiki_talk": 9, "template": 10, "template_talk": 11, "help": 12, "help_talk": 13, "category": 14, "category_talk": 15, "image": 6, "image_talk": 7},
-
wgSiteName="2014.igem.org",
+
wgSiteName="2013.igem.org",
wgCategories=[],
wgCategories=[],
-
wgMWSuggestTemplate="https://2014.igem.org/wiki/api.php?action=opensearch\x26search={searchTerms}\x26namespace={namespaces}\x26suggest",
+
wgMWSuggestTemplate="https://2013.igem.org/wiki/api.php?action=opensearch\x26search={searchTerms}\x26namespace={namespaces}\x26suggest",
-
wgDBname="2014_igem_org",
+
wgDBname="2013_igem_org",
wgSearchNamespaces=[0],
wgSearchNamespaces=[0],
wgMWSuggestMessages=["with suggestions", "no suggestions"],
wgMWSuggestMessages=["with suggestions", "no suggestions"],
wgRestrictionEdit=[],
wgRestrictionEdit=[],
-
wgRestrictionMove=[],
+
wgRestrictionMove=[];
-
wgAjaxWatch={"watchMsg": "Watch", "unwatchMsg": "Unwatch", "watchingMsg": "Watching...", "unwatchingMsg": "Unwatching...", "tooltip-ca-watchMsg": "Add this page to your watchlist", "tooltip-ca-unwatchMsg": "Remove this page from your watchlist"};
+
</script>                 
</script>                 
<script type="text/javascript" src="/wiki/skins/common/wikibits.js?270"><!-- wikibits js --></script>
<script type="text/javascript" src="/wiki/skins/common/wikibits.js?270"><!-- wikibits js --></script>
<!-- Head Scripts -->
<!-- Head Scripts -->
<script src="/wiki/skins/common/ajax.js?270"></script>
<script src="/wiki/skins/common/ajax.js?270"></script>
-
<script src="/wiki/skins/common/ajaxwatch.js?270"></script>
 
<script src="/wiki/skins/common/mwsuggest.js?270"></script>
<script src="/wiki/skins/common/mwsuggest.js?270"></script>
-
<script type="text/javascript" src="/wiki/index.php?title=-&amp;action=raw&amp;smaxage=0&amp;gen=js&amp;useskin=igem"><!-- site js --></script>
+
<script type="text/javascript" src="/wiki/index.php?title=-&amp;action=raw&amp;gen=js&amp;useskin=igem"><!-- site js --></script>
<!-- jQuery Javascript -->
<!-- jQuery Javascript -->
<script type='text/javascript'        src ='/common/jquery-latest.min.js'></script>
<script type='text/javascript'        src ='/common/jquery-latest.min.js'></script>
Line 80: Line 76:
</head>
</head>
-
<body              class="mediawiki  ltr ns-0 ns-subject page-WikitemplateB_project">
+
<body              class="mediawiki  ltr ns-0 ns-subject page-Team_Cornell_notebook">
   <div id="globalWrapper">
   <div id="globalWrapper">
Line 87: Line 83:
    <a href="/Main_Page"
    <a href="/Main_Page"
      title="Main Page">
      title="Main Page">
-
    <img src='https://2014.igem.org/images/wiki.png'>"
+
    <img src='/wiki/skins/common/images/wiki.jpg'>"
    </a>
    </a>
</div>  <!-- end p-logo -->
</div>  <!-- end p-logo -->
Line 94: Line 90:
-
  <div id="menubar" class='left-menu noprint'>
+
  <!--div id="menubar" class='left-menu noprint'>
    <ul>
    <ul>
                    <li  
                    <li  
-
                 class='selected'       ><a href="/WikitemplateB_project">Page              </a></li>
+
                 class='selected'       ><a href="/Team:NTNU_Trondheim/Home">Page              </a></li>
               <li  
               <li  
-
                 class='new'       ><a href="/wiki/index.php?title=Talk:WikitemplateB_project&amp;action=edit&amp;redlink=1">Discussion              </a></li>
+
                 class='new'       ><a href="/wiki/index.php?title=Talk:Team:NTNU_Trondheim/Home&amp;action=edit&amp;redlink=1">Discussion              </a></li>
               <li  
               <li  
-
                     ><a href="/wiki/index.php?title=WikitemplateB_project&amp;action=edit">Edit               </a></li>
+
                     ><a href="/wiki/index.php?title=Team:NTNU_Trondheim/Home&amp;action=edit">View source               </a></li>
               <li  
               <li  
-
                     ><a href="/wiki/index.php?title=WikitemplateB_project&amp;action=history">History               </a></li>
+
                     ><a href="/wiki/index.php?title=Team:NTNU_Trondheim/Home&amp;action=history">History              </a></li>
-
              <li
+
-
                    ><a href="/Special:MovePage/WikitemplateB_project">Move              </a></li>
+
-
              <li
+
-
                    ><a href="/wiki/index.php?title=WikitemplateB_project&amp;action=watch">Watch               </a></li>
+
               <li style='color:white;cursor:default'>teams</li>
               <li style='color:white;cursor:default'>teams</li>
    </ul>
    </ul>
  </div> <!-- end menubar (left) -->
  </div> <!-- end menubar (left) -->
-
<div class="right-menu noprint" id="menubar">
+
 
-
    <ul>
+
 
-
                <li id="pt-userpage"><a href="/User:EivindD" title="Your user page [.]" accesskey="." class="new">EivindD</a></li>                              <li id='pt-preferences'><a href='https://igem.org/User_Information' title='My account'>My account</a></li>
+
 
-
                          <li id="pt-logout"><a href="/wiki/index.php?title=Special:UserLogout&amp;returnto=WikitemplateB_project" title="Log out">Log out</a></li>     </ul>
+
-
</div><!-- end right menubar -->
+
<div id="search-controls" class="noprint">
<div id="search-controls" class="noprint">
Line 128: Line 118:
     <div id="content">
     <div id="content">
<a name="top" id="top"></a>
<a name="top" id="top"></a>
-
<h1 class="firstHeading">WikitemplateB project</h1>
+
<h1 class="firstHeading">Team:NTNU_Trondheim/Home</h1>
<div id="bodyContent">
<div id="bodyContent">
<h3 id="siteSub" class='noprint'>From 2014.igem.org</h3>
<h3 id="siteSub" class='noprint'>From 2014.igem.org</h3>
Line 136: Line 126:
<!-- start content -->
<!-- start content -->
<p>
<p>
-
<head>
+
<head>
-
<style type="text/css">
+
 +
<!--[if lt IE 7]> <html class="no-js lt-ie9 lt-ie8 lt-ie7" lang="en"> <![endif]-->
 +
<!--[if IE 7]>    <html class="no-js lt-ie9 lt-ie8" lang="en"> <![endif]-->
 +
<!--[if IE 8]>    <html class="no-js lt-ie9" lang="en"> <![endif]-->
 +
<!--[if gt IE 8]><!-->
 +
<!-- IE Fix for HTML5 Tags -->
 +
<!--[if lt IE 9]>
 +
<script src="http://html5shiv.googlecode.com/svn/trunk/html5.js"></script>
 +
<![endif]-->
 +
<meta charset="utf-8" />
 +
<meta name="viewport" content="width=device-width" />
 +
<title>NTNU Genetically Engineered Machines</title>
 +
<link rel="stylesheet" type="text/css" href="https://2014.igem.org/Team:NTNU_Trondheim/css/foundation?action=raw&amp;ctype=text/css">
 +
<link rel="stylesheet" type="text/css" href="https://2014.igem.org/Team:NTNU_Trondheim/css/app?action=raw&amp;ctype=text/css">
 +
<style type="text/css">
 +
table#toc {
 +
display:none;
 +
}
-
#contentSub, #footer-box, #catlinks, #search-controls, #p-logo, .printfooter, .visualClear {display: none;} /*-- hides default wiki settings --*/
+
#footer-box {
 +
display:none;
 +
}
 +
</style>
-
.firstHeading {
 
-
width: 975px;
 
-
margin: 0px auto;
 
-
padding-top: 100px;
 
-
margin-bottom: 20px;
 
-
font-family: Georgia, Times, "Times New Roman", serif;
 
-
}
 
-
h1, h2, h3, h4, h5 { font-family: Georgia, Times, "Times New Roman", serif;}
+
<script type="text/javascript" src="https://2012.igem.org/Team:Cornell/javascripts/modernizr.foundation?action=raw&amp;ctype=text/javascript"></script>
 +
<script type="text/javascript" src="https://2014.igem.org/Team:NTNU_Trondheim/javascripts/foundation?action=raw&amp;ctype=text/javascript"></script>
 +
<script type="text/javascript" src="https://2014.igem.org/Team:NTNU_Trondheim/javascripts/app?action=raw&amp;ctype=text/javascript"></script>
 +
<script type="text/javascript" src="https://2014.igem.org/Team:NTNU_Trondheim/javascripts/lightbox?action=raw&amp;ctype=text/javascript"></script>
 +
 +
 +
</head>
 +
<body>
 +
<!-- TOUR TEMPLATE -->
 +
<div class="tour" style="left:0;display:none">
 +
<a id="tourPrev" class="button radius"> Previous<br/>Tour Page </a>
 +
</div>
 +
<div class="tour" style="right:0;display:none">
 +
<a id="tourNext" class="button radius"> Next<br/>Tour Page </a>
 +
</div>
 +
<div class="tour" style="right:0;display:none">
 +
<a id="tourStart" class="button radius"> Start<br/>Tour! </a>
 +
</div>
 +
<!-- HEADER TEMPLATE -->
 +
<div class="row header off-canvas">
 +
<div class="twelve columns centered header no-pad" id="header-wrapper">
 +
<a href="https://2014.igem.org/Team:NTNU_Trondheim">
 +
<div id="logo-wrapper">
 +
<img src="https://static.igem.org/mediawiki/2014/c/ce/NTNU_logo_bar.png" height="6%">
 +
</div> </a>
 +
<ul class="nav-bar">
 +
<li class="pg-home">
 +
<a href="https://2014.igem.org/Team:NTNU_Trondheim">Home</a>
 +
</li>
 +
<li class="has-flyout pg-project pg-project_background pg-project_wetlab pg-project_drylab pg-project_hprac pg-project_safety pg-project_future">
 +
<a href="https://2014.igem.org/Team:NTNU_Trondheim/Project">Project</a>
 +
<a href="https://2014.igem.org/Team:NTNU_Trondheim/Project" class="flyout-toggle"><span> </span></a>
 +
<ul class="flyout">
 +
<li class="pg-project">
 +
<a href="https://2014.igem.org/Team:NTNU_Trondheim/Project">Overview</a>
 +
</li>
 +
<li class="pg-project_background">
 +
<a href="https://2014.igem.org/Team:NTNU_Trondheim/Project/Modelling">Modelling</a>
 +
</li>
 +
<li class="pg-project_background">
 +
<a href="https://2014.igem.org/Team:NTNU_Trondheim/Project/Considerations">Considerations</a>
 +
</li>
 +
<li class="pg-project_safety">
 +
<a href="https://igem.org/Safety/Safety_Form?team_id=1424">Safety</a>
 +
</li>
 +
<li class="pg-project_safety">
 +
<a href="https://2014.igem.org/Team:NTNU_Trondheim/Judging">Judging criteria</a>
 +
</li>
 +
</ul>
 +
</li>
 +
<li class="pg-outreach pg-outreach_presentations pg-outreach_panels pg-outreach_awareness">
 +
<a href="https://2014.igem.org/Team:NTNU_Trondheim/Team">Team</a>
 +
 +
</li>
 +
<li class="pg-notebook pg-notebook_protocols pg-notebook_plasmids">
 +
<a href="https://2014.igem.org/Team:NTNU_Trondheim/Notebook">Notebook</a>
 +
<a href="https://2014.igem.org/Team:NTNU_Trondheim/Notebook" class="flyout-toggle"><span> </span></a>
 +
</li>
 +
<li class="pg-outreach pg-outreach_presentations pg-outreach_panels pg-outreach_awareness">
 +
<a href="https://2014.igem.org/Team:NTNU_Trondheim/Protocols">Protocols</a>
 +
</li>
 +
<li class="has-flyout pg-outreach pg-outreach_presentations pg-outreach_panels pg-outreach_awareness">
 +
<a href="https://2014.igem.org/Team:NTNU_Trondheim/Acknowledgements/Sponsors">Acknowledgements</a>
 +
<a href="https://2014.igem.org/Team:NTNU_Trondheim/Project" class="flyout-toggle"><span> </span></a>
 +
<ul class="flyout">
 +
<li class="pg-project">
 +
<a href="https://2014.igem.org/Team:NTNU_Trondheim/Acknowledgements/Sponsors">Sponsors</a>
 +
</li>
 +
<li class="pg-project_background">
 +
<a href="https://2014.igem.org/Team:NTNU_Trondheim/Acknowledgements/Attributions">Attributions</a>
 +
</li>
 +
</ul>
 +
</li>
 +
<li class="pg-badge">
 +
<div class="badge-wrapper">
 +
<a href="https://2014.igem.org/">
 +
<img style="margin-top:2px;height:50px;width:90px;" src="https://static.igem.org/mediawiki/2014/a/af/IGEM_logo_small.png">
 +
</a>
 +
</div>
 +
</li>
 +
 +
 +
</ul>
 +
<div class="clearfix"></div>
 +
</div>
 +
</div>
-
#top-section { /*-- styling for default menu bar (edit, page, history, etc.) --*/
 
-
background-color: #383838;
 
-
border: 0 none;
 
-
height: 14px;
 
-
z-index: 100;
 
-
top: 0;
 
-
position: fixed;
 
-
width: 975px;
 
-
left: 50%;
 
-
margin-left: -487px;
 
-
}
 
-
#top-section-bar { /*-- styling full width bar which hides behind default menu bar (edit, page, history, etc.) --*/
+
<script type="text/javascript">
-
background-color: #383838;
+
$(window).load(function() {
-
height: 14px;
+
$('li.pg-team_bios').addClass('active');
-
display: block;
+
});
-
z-index: 10;
+
</script>
-
position: fixed;
+
<body>
-
width: 100%;
+
<div class="row">
-
top: 0;
+
<div class="two columns">
-
}
+
<ul class="side-nav">
-
 
+
<li>
-
#menubar a:link, #menubar a:active, #menubar a:visited, #menubar a:hover, #menubar:hover { /*-- styling for default menu bar links (edit, page, history, etc.) --*/
+
<h6>SynECO<sub>2</sub></h6>
-
color: #727272;
+
</li>
-
text-decoration: none;
+
<li class="divider"></li>
-
background-color: transparent;
+
<li>
-
}
+
<a href="#team">Synechocystis conjugation BioBricks</a>
-
 
+
</li>
-
body {
+
<li>
-
background-color: #fff;
+
<a href="#faculty">Carbon fixation BioBrick</a>
-
}
+
</li>
-
 
+
</ul>
-
#globalWrapper, #content { /*-- changes default wiki settings --*/
+
</div>
-
width: 100%;
+
<div class="ten columns team-bios-container">
-
height: 100%;
+
<div id="team" class="row">
-
border: 0px;
+
<div class="twelve columns">
-
background-color: transparent;
+
<h2 class="centered">SynECO<sub>2</sub></h2>
-
margin: 0px;
+
</div>
-
padding: 0px;
+
</div>
-
}
+
<div class="row">
-
 
+
<div class="three columns">
-
html, body, .wrapper { /*-- changes default wiki settings --*/
+
<img src="https://static.igem.org/mediawiki/2014/c/c7/NTNU_atmospheric_co2.png" height="300px">
-
width: 100%;
+
</div>
-
height: 100%;
+
<div class="nine columns">
-
background-color: transparent;
+
<h3>Introduction</h3>
-
}
+
<p>CO<sub>2</sub> emissions have recieved a lot of attention in modern times, due to concerns that high emission levels are facilitating global warming. Consequently, a lot of research is focused on ways of reducing CO<sub>2</sub> emissions from industry, and ways of fixating atmospheric CO<sub>2</sub> at a greater than normal rate.</p>
-
 
+
<p>Our project is attempting to produce a plasmid, which when placed inside photosynthetic bacteria, increases their rate of CO<sub>2</sub> fixation. In order to achieve this, we first need to construct BioBricks that allow inducible expression of non native genes in our chassis; <i>Synechocystis</i> sp. PCC 6803, when assembled.</p>
-
#contentcontainer { /*-- creates container for all content on page --*/
+
</div>
-
font-family: Arial, Helvetica, sans-serif;
+
</div>
-
font-weight: normal;
+
<div class="row">
-
font-size: 14px;
+
<div class="nine columns">
-
color: #414141;
+
<h3>Making a Synechocystis compatible vector</h3>
-
width: 960px;
+
<p>
-
margin-left: auto;
+
In order to start working with <i>Synechocystis</i> sp. PCC 6803, we have to construct a plasmid that will allow insertion of foreign genes into the <i>Synechocystis</i> genome. As <i>Synechocystis</i> does not retain inserted plasmids, transformations must make use of conjugation in order to make the insert part its genome. The flank sequences have been chosen such that they are homologous to a pair of sequences in a neutral site of the <i>Synechocystis</i> genome. When this plasmid is transformed into <i>Synechocystis</i>, the DNA between the two flank sequences is conjugated into its genome.
-
margin-right: auto;
+
</p>
-
background-color: #transparent;
+
<p>
-
margin-top: 0px;
+
The first thing we needed to do was to isolate the flanking sequences from <i>Synechocystis</i>. This was accomplished by use of 'colony PCR', where the genome of <i>Synechocystis</i> was used as a template. We designed primers so that we would only amplify our desired flanking sequences.
-
}
+
</p>
-
 
+
<p>
-
.sidemenu, .sidemenu li {
+
After verifying that our PCR amplified flanking sequences had the right sequence, we wanted to test them. The flanking sequences were tested by ligating them to a vector backbone, with a Kanamycin resistance insert between the two flanking sequences. The resulting plasmid had the following parts, in order:
-
list-style-type: none;
+
<ul>
-
list-style-image: none;
+
<li>Left flank</li>
-
font-family: verdana;
+
<li>Kanamycin resistance</li>
-
text-decoration: none;
+
<li>Right flank</li>
-
color:#000;
+
<li>Backbone</li>
-
font-size: 14px;
+
</ul><br>
-
}
+
This plasmid was transformed into Synechocystis, and the transformed cells were cultured in BG11 growth medium containing kanamycin. Growth of Synechocystis cells in this medium, along with band shift colony PCR (See figure below), confirmed to us that the plasmid had been successfully taken up and integrated into the host genome through homologous recombination.
-
 
+
<img src="https://static.igem.org/mediawiki/2014/c/c0/NTNU_homologous_confirmation.png">
-
.sidemenu li {display: block;}
+
</p>
-
 
+
We wanted to see if transforming the gene <i>glucose oxidase</i> into <i>Synechocystis</i> would lead to an increased rate of carbon fixation pr. growth rate.
-
.sidemenu a {
+
<p>
-
text-decoration:none;
+
To achieve this we constructed a plasmid with eight parts:
-
color: #000;
+
<ul>
-
display: block;
+
<li>"Right flank" sequence</li>
-
width: 180px;
+
<li>Constitutionally active promoter + RBS</li>
-
height: 100%;
+
<li>LacI repressor gene</li>
-
padding: 3px 5px;
+
<li>Kanamycin resistance</li>
-
transition: .25s ease-in-out;
+
<li>LacI inducible promoter + RBS</li>
-
-moz-transition: .25s ease-in-out;
+
<li>Glucose Oxidase</li>
-
-webkit-transition: .25s ease-in-out;
+
<li>"Left flank" sequence</li>
-
}
+
<li>Plasmid backbone</li>
-
 
+
</ul><br>
-
.sidemenu a.greyout { color: #909090; }
+
As growth in <i>Synechocystis</i> is very slow, we did not have time to test this plasmid, and so it will <i>not</i> be submitted as a composite BioBrick.
-
 
+
</p>
-
.sidemenu a:hover {
+
<p>
-
color:#666;
+
Instead, the plasmid was split up into several BioBricks, so that each BioBrick contains one part of the composite plasmid. The BioBricks are as follows:
-
background-color: #d9f5aa;
+
<ul>
-
}
+
<li><a href="http://parts.igem.org/Part:BBa_K1424001">Right flank</a>  (Cloned from <i>Synechocystis</i>)</li>
-
 
+
<li><a href="http://parts.igem.org/Part:BBa_K1424000">Left flank</a> (Cloned from <i>Synechocystis</i>)</li>
-
.sub-sidemenu {font-size: 10px; margin-bottom: 20px; color: #666; display: none;}
+
<li><a href="http://parts.igem.org/Part:BBa_K1424002">LacI inducible promoter + Ribosomal Binding Site </a>(Codon optimized for <i>Synechocystis</i>)</li>
-
.sub-sidemenu a {color: #666;}
+
<li><a href="http://parts.igem.org/Part:BBa_K1424003">Kanamycin resistance gene</a> (Cloned from <i>Synechocystis</i> )</li>
-
 
+
<li><a href="http://parts.igem.org/Part:BBa_K1424004">Glucose Oxidase gene</a> (Synthesized by GenScript, codon optimized for <i>Synechocystis</i>.)</li>
-
div.underconst
+
<li><a href="http://parts.igem.org/Part:BBa_K1424005">Right flank + Kanamycin resistance + Right flank</a></li>
-
{
+
</ul><br>
-
padding:10px;
+
These BioBricks should provide teams with the means to use <i>Synechocystis</i> as a chassis in future iGEM competitions.
-
border-radius:5px;
+
</p>
-
background-color:pink;
+
</div>
-
text-align:center;
+
<div class="three columns">
-
}
+
<img src="" height="200px">
-
 
+
</div>
-
a.anchor{display: block; position: relative; top: -80px; visibility: hidden;}
+
</div>
-
 
+
<div class="row" id="pal">
-
.contentpara {margin-bottom: 30px;}
+
<div class="three columns">
-
 
+
<img src="" height="200px">
-
span.email{font-family: monospace; font-weight: normal;}
+
</div>
-
 
+
<div class="nine columns">
-
fieldset {border: 1px solid #337f53;}
+
<h3>Future efforts</h3>
-
 
+
<p>Our candiate gene for increasing CO<sub>2</sub> the fixation rate is <i>glucose oxidase</i>, which is not originally present in <i>Synechocystis</i>. This gene encodes the enzyme Glucose Oxidase, which essentially reduces the oxygen concentration inside the cell. RuBisCO, the CO<sub>2</sub> fixating enzyme in photosynthetic organisms, has a high affinity for binding O<sub>2</sub>, which can interfere with CO<sub>2</sub> binding. Reducing O<sub>2</sub> concentrations could therefore lead to an increased rate of CO<sub>2</sub> fixation in <i>Synechocystis</i>.</p>
-
.greyout {color: #A0A0A0;}
+
<p>Future efforts should focus on transforming the composite plasmid containing all 6 BioBricks into Synechocystis, and testing the carbon fixation rate of the resulting transformants. Such testing requires specialized equipment that is able to measure minute differences in the partial pressure of CO<sub>2<sub>.</p>  
-
.highlightme {background-color: #FFFF00;}
+
-
 
+
-
#alertContainer {border: 2px solid red; margin-bottom: 10px; width: 936px;}
+
-
#annContainer {margin-left: 8px;}
+
-
#newsContainer {border: 1px solid #ccc;}
+
-
 
+
-
.newsTitle {
+
-
display: block;
+
-
color: #414141;
+
-
font-size: 25px; 
+
-
        font-family: Georgia, Times, "Times New Roman", serif;
+
-
padding: 10px 15px 5px 10px;
+
-
border-bottom: 1px solid #ccc;
+
-
margin-bottom: 0px;
+
-
}
+
-
 
+
-
.newsItem {
+
-
border-bottom: 1px solid #ccc;
+
-
display: block;
+
-
padding: 5px 15px 0px 10px;
+
-
margin-bottom: 0px;
+
-
}
+
-
 
+
-
.newsItem h3 {
+
-
width: auto;
+
-
display: inline;
+
-
font-size: 14px;
+
-
font-family: Arial, Helvetica, sans-serif;
+
-
padding: 0px;
+
-
margin: 0px;
+
-
}
+
-
 
+
-
.newsItem img {
+
-
float: right;
+
-
clear: right;
+
-
width: 80px;
+
-
padding-left: 15px;
+
-
display: inline-block;
+
-
}
+
-
 
+
-
.newsItem .newsDate {
+
-
font-style: italic;
+
-
font-size: 14px;
+
-
display: inline-block;
+
-
color: #999;
+
-
float: right;
+
-
padding: 0px 0px 10px 15px;
+
-
}
+
-
 
+
-
.newsItem p, .newsItem ul, .newsItem li {
+
-
padding: 0px;
+
-
margin-left: 20px;
+
-
font-size: 12px;
+
-
line-height: 1.2;
+
-
}
+
-
 
+
-
.newsItem p {margin-top: 10px;}
+
-
 
+
-
.annItem {
+
-
border: 2px solid #49b677;
+
-
display: block;
+
-
padding: 0px 15px 10px 10px;
+
-
margin-bottom: 18px;
+
-
position: relative;
+
-
height: 160px;
+
-
vertical-align: middle;
+
-
}
+
-
 
+
-
.annItem .annCentered {
+
-
display: table-cell;  
+
-
vertical-align: middle;
+
-
height: 160px;
+
-
}
+
-
 
+
-
.annCentered h3 {
+
-
padding: 0px;
+
-
display: block;
+
-
margin-left: auto;
+
-
margin-right: auto;
+
-
text-align: center;
+
-
}
+
-
 
+
-
.annItem ul, .annItem li {padding: 0px; margin: 0px;}
+
-
.annItem li {list-style: none; margin: 5px}
+
-
 
+
-
 
+
-
 
+
-
</style>
+
-
</style>
+
-
 
+
-
 
+
-
 
+
-
 
+
-
 
+
-
<style>
+
-
#contentSub, #footer-box, #catlinks, #search-controls, #p-logo, .printfooter, .firstHeading,.visualClear {display: none;} /*-- hides default wiki settings --*/
+
-
</style>
+
-
 
+
-
<!--  here ends the section that changes the default wiki template to a white full width background -->
+
-
 
+
-
 
+
-
<!-- beginning of your page -->
+
-
 
+
-
<!-- menu -->
+
-
<table id="menu" width="100%"  cellspacing="0" height="135px">
+
-
<tr>
+
-
<td  bgColor="#FFFFFF"></td>
+
-
<td valign="bottom"  width="975px" align="center" bgColor="#FFFFFF">
+
-
 
+
-
<table align="center">
+
-
<tr height="50px">
+
-
<td width="162px" align="center" onMouseOver="this.bgColor='#FEE5AD'" onMouseOut="this.bgColor='#A1DBB2'" bgColor=#A1DBB2>  
+
-
<a href="https://2014.igem.org/Team:NTNU_Trondheim"style="text-decoration:none;color:#1C140D">HOME </a> </td>  
+
-
 
+
-
<td width="162px" align="center" onMouseOver="this.bgColor='#FEE5AD'" onMouseOut="this.bgColor='#A1DBB2'" bgColor=#A1DBB2>  
+
-
<a href="https://2014.igem.org/Team:NTNU_Trondheim/Project" style="text-decoration:none;color:#1C140D">PROJECT</a> </td>
+
-
 
+
-
<td width="162px" align="center" onMouseOver="this.bgColor='#FEE5AD'" onMouseOut="this.bgColor='#A1DBB2'" bgColor=#A1DBB2>  
+
-
<a href="https://2014.igem.org/Team:NTNU_Trondheim/Parts" style="text-decoration:none;color:#1C140D">PARTS</a></td>  
+
-
 
+
-
<td width="162px" align="center" onMouseOver="this.bgColor='#FEE5AD'" onMouseOut="this.bgColor='#A1DBB2'" bgColor=#A1DBB2>  
+
-
<a href="https://2014.igem.org/Team:NTNU_Trondheim/Team" style="text-decoration:none;color:#1C140D">TEAM </a> </td>
+
-
 
+
-
<td width="162px" align="center" onMouseOver="this.bgColor='#FEE5AD'" onMouseOut="this.bgColor='#A1DBB2'" bgColor=#A1DBB2>
+
-
<a href="https://2014.igem.org/Team:NTNU_Trondheim/Notebook" style="text-decoration:none;color:#1C140D">NOTEBOOK </a> </td>
+
-
 
+
-
<td width="162px" align="center" onMouseOver="this.bgColor='#FEE5AD'" onMouseOut="this.bgColor='#A1DBB2'" bgColor=#A1DBB2>
+
-
<a href="https://2014.igem.org/Team:NTNU_Trondheim/Safety" style="text-decoration:none;color:#1C140D">SAFETY </a></td>
+
-
</tr>
+
-
 
+
-
<tr> <td colspan="5"></td>  
+
-
<td align="center" width="162px"><a href="https://igem.org/Main_Page"> <img src="https://static.igem.org/mediawiki/igem.org/6/60/Igemlogo_300px.png" width="55px"></a></td>  
+
-
</tr>
+
-
</table>
+
-
 
+
-
</td>
+
-
<td  bgColor="#FFFFFF"></td>
+
-
</tr>
+
-
</table>
+
-
<!-- end of menu -->
+
-
 
+
-
 
+
-
 
+
-
 
+
-
<!-- Page content begins -->
+
-
 
+
-
 
+
-
<!-- first block of three columns -->
+
-
<table width="100%"  cellspacing="0" height="230px">
+
-
<tr><td  bgColor="#FEE5AD"></td> <td colspan="3" width="975px" bgColor="#FEE5AD" align="center"> <h3>Team Example's Project name! </h3></td> <td  bgColor="#FEE5AD"></td> </tr>
+
-
 
+
-
<tr>
+
-
<td  bgColor="#FFFFFF"></td>
+
-
 
+
-
<td width="435px"  height="400px"  bgColor="#FFFFFF" >
+
-
<h3>Project Introduction</h3>
+
-
<p>CO<sub>2</sub> emissions have recieved a lot of attention in modern times, due to concerns that high emission levels are facilitating global warming. Consequently, a lot of research is focused on ways of reducing CO<sub>2</sub> emissions from industry, and ways of fixating atmospheric CO<sub>2</sub> at a greater than normal rate.</p>
+
-
 
+
-
<p>Our project is attempting to produce a BioBrick, which when placed inside photosynthetic bacteria, increases their rate of CO<sub>2</sub> fixation. In order to achieve this, we first need to construct a BioBrick that allows inducible expression of non native genes in our chassis; <a href="http://www.genome.jp/kegg-bin/show_organism?org=syn"><i>Synechocystis</i> sp. PCC 6803</a></p>
+
-
</td>
+
-
 
+
-
<td width="50px"></td>
+
-
<td width="435px" bgColor="#FFFFFF">
+
-
<h4>Results</h4>
+
-
<ul>
+
-
<li>Result 1 - Lorem ipsum ad his scripta blandit partiendo, eum fastidii accumsan euripidis in, eum liber hendrerit an.</li>
+
-
<li>Result 2 - Lorem ipsum ad his scripta blandit partiendo, eum fastidii accumsan euripidis in, eum liber hendrerit an.</li>  
+
-
<li>Result 3 - Lorem ipsum ad his scripta blandit partiendo, eum fastidii accumsan euripidis in, eum liber hendrerit an.</li>
+
-
 
+
-
</ul>
+
-
</td>
+
-
 
+
-
<td  bgColor="#FFFFFF"></td>
+
-
</tr>
+
-
</table>
+
-
<!-- end of first block -->
+
-
 
+
-
<!-- navigation for project page-->
+
-
<table width="100%" align="center" >
+
-
 
+
-
<tr>
+
-
<td></td>
+
-
<td align="center" width="10%" height ="35px"
+
-
onMouseOver="this.bgColor='#A1DBB2'" onMouseOut="this.bgColor='#FEE5AD'" bgColor=#FEE5AD>
+
-
<a href="#reporter" style="text-decoration:none;color:#000000">Reporter BioBrick </a> </td>
+
-
</td>
+
-
 
+
-
<td align="center" width="10%" height ="35px"
+
-
onMouseOver="this.bgColor='#A1DBB2'" onMouseOut="this.bgColor='#FEE5AD'" bgColor=#FEE5AD> 
+
-
<a href="#co2" style="text-decoration:none;color:#000000">CO<sub>2</sub> fixation BioBrick </a> </td>
+
-
</td>
+
-
 
+
-
<td align="center" width="10%" height ="35px"
+
-
onMouseOver="this.bgColor='#A1DBB2'" onMouseOut="this.bgColor='#FEE5AD'" bgColor=#FEE5AD>
+
-
<a href="#testing" style="text-decoration:none;color:#000000">Testing CO<sub>2</sub> fixation rate </a> </td>
+
-
</td>
+
-
 
+
-
<td></td>
+
-
</tr>
+
-
</table>
+
-
<!-- end of navigation page -->
+
-
 
+
-
 
+
-
 
+
-
<!-- abstract-->
+
-
<table width="100%"  cellspacing="0" height="275px">
+
-
 
+
-
<tr>
+
-
<td  bgColor="#FFFFFF"></td>
+
-
 
+
-
<td width="450px"  bgColor="#FFFFFF">
+
-
<h4 id="reporter"> Reporter BioBrick</h4>
+
-
<p>The first stage of this project consists of creating a reporter BioBrick in order to confirm that we have are able to induce the expression of foreign genes in <a href="http://www.genome.jp/kegg-bin/show_organism?org=syn"><i>Synechocystis</i> sp. PCC 6803</a>. This reporter BioBrick makes use of several other BioBricks, such as <a href="http://parts.igem.org/Part:BBa_J23101">BBa_J23101</a>, <a href="http://parts.igem.org/Part:BBa_B0034">BBa_B0034</a> and <a href="http://parts.igem.org/Part:BBa_C0012">BBa_C0012</a>.</p>
+
-
 
+
-
<p>Once the reporter plasmid has been successfully constructed, we will verify that it is working as intended by inducing the expression of a fluorescent protein contained on the plasmid. Once induced, the colonies of cells transformed with this plasmid should exhibit a color that is determined by the fluorescent protein. </p>
+
-
 
+
-
<p>The plasmid is transformed into <i>Synechocystis</i> sp. PCC 6803 by homologous recombination. Contained on the plasmid are left- and right flanking sequences obtained from the <i>Synechocystis</i> genome. These flanking sequences undergo homologous with their sister sequences in the genome of <i>Synechocystis</i>, integrating the plasmid sequence into the genome. </p>
+
-
</td>
+
-
 
+
-
<td width="20px"></td>
+
-
<td width="450px" bgColor="#FFFFFF">
+
-
<img src="https://static.igem.org/mediawiki/2014/4/41/NTNU_Trondheim_reporter_plasmid.png" width="400px">
+
-
<p> The reporter plasmid will use a fluorescent protein placed under the control of an indicuble promoter. This will allow us to easily verify that the inducible expression system works as intended inside <i>Synechocystis</i> sp. PCC 6803.</p>
+
-
</td>
+
-
 
+
-
<td  bgColor="#FFFFFF"></td>
+
-
</tr>
+
-
 
+
-
<tr height="100px"></tr>
+
-
 
+
-
<tr>
+
-
<td  bgColor="#FFFFFF"></td>
+
-
 
+
-
<td width="450px" bgColor="#FFFFFF">
+
-
<h4 id="fixating"> CO<sub>2</sub> fixating BioBrick</h4>
+
-
<p>After verifying that the expression system is working as intended in <i>Synechocystis</i>, we will replace the fluorescent protein gene with one that is predicted to increase the rate of carbon fixation in the organism. The way we will idenfity this gene is by use of metabolic modelling. Once a candidate gene has been detected, it will be cloned and transformed into <i>Synechocystis</i>.</p>
+
-
</td>
+
-
 
+
-
<td width="20px"></td>
+
-
<td width="450px" bgColor="#FFFFFF">
+
-
<img src="" width="400px">
+
-
<p> </p>
+
-
</td>
+
-
 
+
-
<td  bgColor="#FFFFFF"></td>
+
-
</tr>
+
-
 
+
-
</table>
+
-
<!-- end of first block -->
+
 +
<p>We had plans of collaborating with <a href="http://www.sintef.no/home/Environment/CO2-fangst-og-handtering/"> SINTEF CO<sub>2</sub>-capture and storage</a>, which were kind enough to offer us the use of their facilities and equipment. Doing our planned experiments in their facilities would have provided us with evidence for the effect of <i>glucose oxidase</i> on <i>Synechocystis'</i> carbon fixation rate.</p>
 +
</div>
 +
</div>
 +
</div>
 +
</div>
 +
</body>
</p>
</p>
 +
<body>
 +
<div id="footerdiv">
 +
<center>
 +
<a href="http://www.genscript.com"><img src="https://static.igem.org/mediawiki/2014/0/00/NTNU_genscript_logo_small.png" width ="200"></a>
 +
<a href="http://www.skretting.no"><img src="https://static.igem.org/mediawiki/2014/7/74/Skretting_logo.png" width="200"></a>
 +
<a href="http://www.sparebank1.no/smn"><img src="https://static.igem.org/mediawiki/2014/9/98/SpareBank1_RGB_nett.jpg" width="200"></a>
 +
<a href="http://www.geneious.com/"><img src="https://static.igem.org/mediawiki/2014/d/d6/Genious_logo.png" width="200px"></a>
 +
<a href="http://www.enova.no/"><img src="https://static.igem.org/mediawiki/2014/2/2d/NTNU_enova_small.png" width="200px"></a>
 +
</center>
 +
</div>
 +
</body>
<!--  
<!--  
NewPP limit report
NewPP limit report
Preprocessor node count: 9/1000000
Preprocessor node count: 9/1000000
-
Post-expand include size: 41/2097152 bytes
+
Post-expand include size: 42/2097152 bytes
Template argument size: 0/2097152 bytes
Template argument size: 0/2097152 bytes
Expensive parser function count: 0/100
Expensive parser function count: 0/100
-->
-->
-
<!-- Saved in parser cache with key 2014_igem_org:pcache:idhash:1048-0!1!0!!en!2 and timestamp 20140625093431 -->
+
<!-- Saved in parser cache with key 2013_igem_org:pcache:idhash:11247-0!1!0!!en!2!edit=0 and timestamp 20140805122150 -->
<div class="printfooter">
<div class="printfooter">
-
Retrieved from "<a href="https://2014.igem.org/WikitemplateB_project">https://2014.igem.org/WikitemplateB_project</a>"</div>
+
Retrieved from "<a href="https://2013.igem.org/Team:Cornell/team/bios">https://2013.igem.org/Team:Cornell/team/bios</a>"</div>
<div id="catlinks"><div id='catlinks' class='catlinks catlinks-allhidden'></div></div> <!-- end content -->
<div id="catlinks"><div id='catlinks' class='catlinks catlinks-allhidden'></div></div> <!-- end content -->
<div class="visualClear"></div>
<div class="visualClear"></div>
Line 536: Line 381:
  <!-- Recentchanges is not handles well DEBUG -->
  <!-- Recentchanges is not handles well DEBUG -->
    <li id="t-recentchanges"><a href="/Special:RecentChanges"
    <li id="t-recentchanges"><a href="/Special:RecentChanges"
-
      title='Recent changes'>Recent changes</a></li>
+
      title='Recent changes'>Recent changesa></li>
-
    <li id="t-whatlinkshere"><a href="/Special:WhatLinksHere/WikitemplateB_project"
+
    <li id="t-whatlinkshere"><a href="/Special:WhatLinksHere/Team:Cornell/team/bios"
      title="List of all wiki pages that link here [j]" accesskey="j">What links here</a></li>
      title="List of all wiki pages that link here [j]" accesskey="j">What links here</a></li>
-
                         <li id="t-recentchangeslinked"><a href="/Special:RecentChangesLinked/WikitemplateB_project"
+
                         <li id="t-recentchangeslinked"><a href="/Special:RecentChangesLinked/Team:Cornell/team/bios"
                           title="Recent changes in pages linked from this page [k]" accesskey="k">Related changes</a></li>
                           title="Recent changes in pages linked from this page [k]" accesskey="k">Related changes</a></li>
-
                <li id="t-upload"><a href="/Special:Upload"
 
-
                  title="Upload files [u]" accesskey="u">Upload file</a>
 
-
                </li>
 
                <li id="t-specialpages"><a href="/Special:SpecialPages"
                <li id="t-specialpages"><a href="/Special:SpecialPages"
                  title="List of all special pages [q]" accesskey="q">Special pages</a>
                  title="List of all special pages [q]" accesskey="q">Special pages</a>
Line 558: Line 400:
    <ul id="f-list">
    <ul id="f-list">
-
            <li id="t-print"><a href="/wiki/index.php?title=WikitemplateB_project&amp;printable=yes"
+
            <li id="t-print"><a href="/wiki/index.php?title=Team:Cornell/team/bios&amp;printable=yes"
              title="Printable version of this page [p]" accesskey="p">Printable version</a>
              title="Printable version of this page [p]" accesskey="p">Printable version</a>
            </li>
            </li>
-
            <li id="t-permalink"><a href="/wiki/index.php?title=WikitemplateB_project&amp;oldid=4634"
+
            <li id="t-permalink"><a href="/wiki/index.php?title=Team:Cornell/team/bios&amp;oldid=362783"
              title="Permanent link to this revision of the page">Permanent link</a>
              title="Permanent link to this revision of the page">Permanent link</a>
            </li>
            </li>
-
          <li id="privacy"><a href="/2014.igem.org:Privacy_policy" title="2014.igem.org:Privacy policy">Privacy policy</a></li>
+
          <li id="privacy"><a href="/2013.igem.org:Privacy_policy" title="2013.igem.org:Privacy policy">Privacy policy</a></li>
-
        <li id="disclaimer"><a href="/2014.igem.org:General_disclaimer" title="2014.igem.org:General disclaimer">Disclaimers</a></li>
+
        <li id="disclaimer"><a href="/2013.igem.org:General_disclaimer" title="2013.igem.org:General disclaimer">Disclaimers</a></li>
    </ul>
    </ul>
</div> <!-- close footer -->
</div> <!-- close footer -->
Line 575: Line 417:
<script>if (window.runOnloadHook) runOnloadHook();</script>
<script>if (window.runOnloadHook) runOnloadHook();</script>
</div>
</div>
-
<!-- Served in 0.201 secs. --></body>
+
<!-- Served in 0.163 secs. --></body>
</html>
</html>

Latest revision as of 22:08, 17 October 2014

Team:NTNU_Trondheim/notebook - 2014.igem.org

 

Team:NTNU_Trondheim/Home

From 2014.igem.org

NTNU Genetically Engineered Machines

SynECO2

Introduction

CO2 emissions have recieved a lot of attention in modern times, due to concerns that high emission levels are facilitating global warming. Consequently, a lot of research is focused on ways of reducing CO2 emissions from industry, and ways of fixating atmospheric CO2 at a greater than normal rate.

Our project is attempting to produce a plasmid, which when placed inside photosynthetic bacteria, increases their rate of CO2 fixation. In order to achieve this, we first need to construct BioBricks that allow inducible expression of non native genes in our chassis; Synechocystis sp. PCC 6803, when assembled.

Making a Synechocystis compatible vector

In order to start working with Synechocystis sp. PCC 6803, we have to construct a plasmid that will allow insertion of foreign genes into the Synechocystis genome. As Synechocystis does not retain inserted plasmids, transformations must make use of conjugation in order to make the insert part its genome. The flank sequences have been chosen such that they are homologous to a pair of sequences in a neutral site of the Synechocystis genome. When this plasmid is transformed into Synechocystis, the DNA between the two flank sequences is conjugated into its genome.

The first thing we needed to do was to isolate the flanking sequences from Synechocystis. This was accomplished by use of 'colony PCR', where the genome of Synechocystis was used as a template. We designed primers so that we would only amplify our desired flanking sequences.

After verifying that our PCR amplified flanking sequences had the right sequence, we wanted to test them. The flanking sequences were tested by ligating them to a vector backbone, with a Kanamycin resistance insert between the two flanking sequences. The resulting plasmid had the following parts, in order:

  • Left flank
  • Kanamycin resistance
  • Right flank
  • Backbone

This plasmid was transformed into Synechocystis, and the transformed cells were cultured in BG11 growth medium containing kanamycin. Growth of Synechocystis cells in this medium, along with band shift colony PCR (See figure below), confirmed to us that the plasmid had been successfully taken up and integrated into the host genome through homologous recombination.

We wanted to see if transforming the gene glucose oxidase into Synechocystis would lead to an increased rate of carbon fixation pr. growth rate.

To achieve this we constructed a plasmid with eight parts:

  • "Right flank" sequence
  • Constitutionally active promoter + RBS
  • LacI repressor gene
  • Kanamycin resistance
  • LacI inducible promoter + RBS
  • Glucose Oxidase
  • "Left flank" sequence
  • Plasmid backbone

As growth in Synechocystis is very slow, we did not have time to test this plasmid, and so it will not be submitted as a composite BioBrick.

Instead, the plasmid was split up into several BioBricks, so that each BioBrick contains one part of the composite plasmid. The BioBricks are as follows:


These BioBricks should provide teams with the means to use Synechocystis as a chassis in future iGEM competitions.

Future efforts

Our candiate gene for increasing CO2 the fixation rate is glucose oxidase, which is not originally present in Synechocystis. This gene encodes the enzyme Glucose Oxidase, which essentially reduces the oxygen concentration inside the cell. RuBisCO, the CO2 fixating enzyme in photosynthetic organisms, has a high affinity for binding O2, which can interfere with CO2 binding. Reducing O2 concentrations could therefore lead to an increased rate of CO2 fixation in Synechocystis.

Future efforts should focus on transforming the composite plasmid containing all 6 BioBricks into Synechocystis, and testing the carbon fixation rate of the resulting transformants. Such testing requires specialized equipment that is able to measure minute differences in the partial pressure of CO2.

We had plans of collaborating with SINTEF CO2-capture and storage, which were kind enough to offer us the use of their facilities and equipment. Doing our planned experiments in their facilities would have provided us with evidence for the effect of glucose oxidase on Synechocystis' carbon fixation rate.