Team:NTNU Trondheim/template
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- | class='selected' ><a href="/Team: | + | class='selected' ><a href="/Team:NTNU_Trondheim/Home">Page </a></li> |
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- | class='new' ><a href="/wiki/index.php?title=Talk:Team: | + | class='new' ><a href="/wiki/index.php?title=Talk:Team:NTNU_Trondheim/Home&action=edit&redlink=1">Discussion </a></li> |
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- | ><a href="/wiki/index.php?title=Team: | + | ><a href="/wiki/index.php?title=Team:NTNU_Trondheim/Home&action=history">History </a></li> |
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<input type='submit' name="fulltext" class="searchButton" id="mw-searchButton" value="Search" title="Search the pages for this text" /> | <input type='submit' name="fulltext" class="searchButton" id="mw-searchButton" value="Search" title="Search the pages for this text" /> | ||
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<div class="two columns"> | <div class="two columns"> | ||
- | <ul class="side-nav | + | <ul class="side-nav"> |
- | < | + | <li> |
- | + | <h6>Project description</h6> | |
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- | <li | + | <li class="divider"></li> |
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+ | <a href="#team">Synechocystis conjugation BioBricks</a> | ||
</li> | </li> | ||
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- | <a | + | <a href="#faculty">Carbon fixation BioBrick</a> |
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- | <div | + | <div id="team" class="row"> |
<div class="twelve columns"> | <div class="twelve columns"> | ||
- | <h2 class="centered"> | + | <h2 class="centered">Project Description</h2> |
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- | <div class="row | + | <div class="three columns"> |
- | + | <img src="https://static.igem.org/mediawiki/2014/c/c7/NTNU_atmospheric_co2.png" height="300px"> | |
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- | <div class=" | + | <div class="nine columns"> |
- | < | + | <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 BioBrick, which when placed inside photosynthetic bacteria, increases their rate of CO2 fixation. In order to achieve this, we first need to construct a BioBrick that allows inducible expression of non native genes in our chassis; Synechocystis sp. PCC 6803.</p> | |
- | < | + | |
</div> | </div> | ||
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- | + | <div class="row" id="ronja"> | |
- | < | + | <div class="nine columns"> |
- | + | <h3>Making a Synechocystis compatible vector</h3> | |
- | + | <p> | |
+ | In order to start working with <i>Synechocystis</i>, 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. | ||
+ | </p> | ||
+ | <p> | ||
+ | To achieve this we have constructed a plasmid with eight parts: | ||
+ | <ul> | ||
+ | <li>"Right flank" sequence</li> | ||
+ | <li>Constitutionally active promoter + RBS</li> | ||
+ | <li>LacI repressor gene</li> | ||
+ | <li>Kanamycin resistance</li> | ||
+ | <li>LacI inducible promoter + RBS</li> | ||
+ | <li>Red fluorescent protein</li> | ||
+ | <li>"Left flank" sequence</li> | ||
+ | <li>Plasmid backbone</li> | ||
+ | </ul><br> | ||
+ | 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. By growing cultures in a selection medium containing Kanamycin, the insert is kept inside the genome, allowing us to express foreign genes inside the organism. | ||
+ | </p> | ||
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- | <div class=" | + | <div class="three columns"> |
- | <div class=" | + | <img src="" height="200px"> |
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- | + | <div class="nine columns"> | |
- | + | <h3>Increasing the CO<sub>2</sub> fixation rate</h3> | |
- | + | <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 Synechocystis.</p> | |
- | + | <p>One candiate gene 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> | |
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+ | </div> | ||
</body> | </body> | ||
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+ | <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> | ||
+ | </center> | ||
+ | </div> | ||
+ | </body> | ||
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NewPP limit report | NewPP limit report | ||
- | Preprocessor node count: | + | Preprocessor node count: 9/1000000 |
- | Post-expand include size: | + | Post-expand include size: 42/2097152 bytes |
- | Template argument size: | + | Template argument size: 0/2097152 bytes |
Expensive parser function count: 0/100 | Expensive parser function count: 0/100 | ||
--> | --> | ||
- | <!-- Saved in parser cache with key 2013_igem_org:pcache:idhash: | + | <!-- 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://2013.igem.org/Team:Cornell/ | + | Retrieved from "<a href="https://2013.igem.org/Team:Cornell/team/bios">https://2013.igem.org/Team:Cornell/team/bios</a>"</div> |
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- | title='Recent changes'>Recent | + | title='Recent changes'>Recent changesa></li> |
- | <li id="t-whatlinkshere"><a href="/Special:WhatLinksHere/Team:Cornell/ | + | <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/Team:Cornell/ | + | <li id="t-recentchangeslinked"><a href="/Special:RecentChangesLinked/Team:Cornell/team/bios" |
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title="Printable version of this page [p]" accesskey="p">Printable version</a> | title="Printable version of this page [p]" accesskey="p">Printable version</a> | ||
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- | <li id="t-permalink"><a href="/wiki/index.php?title=Team:Cornell/ | + | <li id="t-permalink"><a href="/wiki/index.php?title=Team:Cornell/team/bios&oldid=362783" |
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Latest revision as of 10:18, 15 August 2014
Team:NTNU_Trondheim/Home
From 2014.igem.org
-
Project description
- Synechocystis conjugation BioBricks
- Carbon fixation BioBrick
Project Description
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 BioBrick, which when placed inside photosynthetic bacteria, increases their rate of CO2 fixation. In order to achieve this, we first need to construct a BioBrick that allows inducible expression of non native genes in our chassis; Synechocystis sp. PCC 6803.
Making a Synechocystis compatible vector
In order to start working with Synechocystis, 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.
To achieve this we have constructed a plasmid with eight parts:
- "Right flank" sequence
- Constitutionally active promoter + RBS
- LacI repressor gene
- Kanamycin resistance
- LacI inducible promoter + RBS
- Red fluorescent protein
- "Left flank" sequence
- Plasmid backbone
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. By growing cultures in a selection medium containing Kanamycin, the insert is kept inside the genome, allowing us to express foreign genes inside the organism.
Increasing the CO2 fixation rate
After verifying that the expression system is working as intended in Synechocystis, 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 Synechocystis.
One candiate gene 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.