Team:CSU Fort Collins

From 2014.igem.org

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     <h1>Welcome to CSU Fort Collin's 2014 Wiki</h1>
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     <h1>Table of Contents</h1>
    
    
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       <h3>Our Project</h3>
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      <h2>1</h2>
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      <h2>Protocols</h2>
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      <span class='index'>01</span>
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      <a href="/Team:CSU_Fort_Collins/Notebook/Protocols=Gibson" class='month-group'>Gibson Assembly</a><br>
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       <span class='index'>02</span>
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      <a href="/Team:CSU_Fort_Collins/Notebook/Protocols=Cloning" class='month-group'>Cloning Genes</a><br>
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      <span class="index">03</span>
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      <a href="/Team:CSU_Fort_Collins/Notebook/Protocols=Miniprep" class='month-group'>Plasmid Miniprep</a><br>
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      <span class="index">04</span>
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      <a href="/Team:CSU_Fort_Collins/Notebook/Protocols=Isolation" class='month-group'>Yeast DNA Extraction</a><br>
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      <span class="index">05</span>
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      <a href="/Team:CSU_Fort_Collins/Notebook/Protocols=Gel" class='month-group'>Gel Electrophoresis</a><br>
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      <span class="index">06</span>
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      <a href="/Team:CSU_Fort_Collins/Notebook/Protocols=Purify" class='month-group'>PCR Product Purification</a><br>
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       Approximately 3 billion gallons per year of used frying oil are produced in the U.S. alone. While some recycling efforts are put forth to turn used frying oil into biodiesel and some companies are working on more effective ways to recycle the frying oil, the iGEM team at Colorado State University is working to turn spent frying oil into a high value product. There are four major components to this year’s project including; the breakdown of frying oil, a biosensor for detecting the breakdown of frying oil, production of a high value product, and a kill switch to kill the bacteria if they were to be released into the environment. Our team aims to put each of these components into Escherichia coli. 
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       <h2>2</h2>
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In order to detect that the process is working we have developed a biosensor that will detect the breakdown of Acyl-CoA, a byproduct of the breakdown pathway of lipids, in E. coli. Our biosensor is being constructed by inserting a novel promoter in front of DNA expressing a Green Fluorescent Protein (GFP). This promoter is activated by Acyl-CoA, a byproduct of the breakdown pathway of lipids in E. coli. This will result in E. coli expressing GFP when successful breakdown is occurring.
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      <h2>Daily Notes</h2>
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      <h4>Biosensor</h4>
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In order to break down the frying oil we will be taking advantage of the cell’s natural ability to break down fatty acids for use in the Kreb’s cycle. We will be upregulating the limiting enzymes that aid in the breakdown of fatty acids in order to have the cell produce more Acetyl-CoA.
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      <span class="index">07</span>
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This Acetyl-CoA can then be used in the mevalonate pathway. This pathway is common to most plants as well as yeast. We will be taking the genes from yeast and constructing two operons in order to produce isopentenyl pyrophosphate (IPP). While both IPP and dimethylallyl pyrophosphate (DMAPP) are both naturally produced in E. coli  through the non-mevalonate pathway , the mevalonate pathway provides a more efficient route from acetyl-CoA to terpenoid production than the alternative. Our high-value product will be a terpenoid that has yet to be determined.
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      <span class='month-group'>
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        <a href='#'>June</a>
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For our kill switch, we will be using a gene named Killer Red which is a mutant of hydrozoan chromoprotein anm2CP from Anthomedusae sp. DC-2005 . Our team plans to insert this gene into our construct and by doing so, it will provide a fail-safe in the unlikely event of our E. coli escaping into the environment. The Killer Red gene has a repressible promoter in which tryptophan acts as the repressor. There will be tryptophan present while the cell is used to break down the frying oil, but is not highly present in the environment. Without the presence of tryptophan the Killer Red gene will be activated and white light will produce reactive oxygen species within the cell, causing cell death.
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        : PAR, PLR, and PFL Assembly<br>
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Our biosensor is being constructed by inserting a novel promoter in front of DNA expressing a Green Fluorescent Protein (GFP). This promoter is activated by Acyl-CoA, a byproduct of the breakdown pathway of lipids in E. coli. This will result in E. coli expressing GFP when successful breakdown is occurring.
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      <span class="index">08</span>
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      <span class='month-group'>
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        <a href='#'>July</a>
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      : PLR, PFL, and PFL2 Assembly<br>
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      <span class="index">09</span>
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      <span class='month-group'>
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      <a href='#'>August</a>
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      <span class="index">10</span>
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      <span class='month-group'>
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        <a href='#'>September</a>
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      : Hungry Hungry Hipsters<br>
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      <h4>Breakdown</h4>
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      <span class="index">11</span>
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      <span class='month-group'>
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        <a href='#'>July</a>
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      : Lac Promoter and FadD Assembly<br>
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      <span class="index">12</span>
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      <span class="month-group">
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        <a href='#' class='month-group'>August</a>
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      <span class="index">13</span>
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      <span class='montth-group'>
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        <a href='#' class='month-group'>September</a>
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      : Hungry Hungry Hipsters<br>
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      <h4>High-Value Product</h4>
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      <span class="index">14</span>
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      <span class='month-group'>
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        <a href='#' class='month-group'>June</a>
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      </span>
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      : pMBI Assembly<br>
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      <span class="index">15</span>
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      <span class="month-group"><a href='#' class='month-group'>July</a>
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      : pMBI and pMevT Assembly<br>
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      <span class="index">16</span> <a href='#' class='month-group'>August</a>: Troubleshooting Gibson Assembly<br>
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      <span class="index">17</span> <a href='#' class='month-group'>September</a>: Hungry Hungry Hipsters<br>
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      <h4>Kill Switch</h4>
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      <span class="index">18</span> <a href='#' class='month-group'>July</a>: Hello this is a Description<br>
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      <span class="index">19</span> <a href='#' class='month-group'>August</a>: Lorem ipsum dolor sit amet<br>
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      <span class="index">20</span> <a href='#' class='month-group'>September</a>: Hungry Hungry Hipsters<br>
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Revision as of 15:23, 7 October 2014

CSU iGEM 2014

Table of Contents

1

Protocols

01 Gibson Assembly
02 Cloning Genes
03 Plasmid Miniprep
04 Yeast DNA Extraction
05 Gel Electrophoresis
06 PCR Product Purification

2

Daily Notes

Biosensor

07 June : PAR, PLR, and PFL Assembly
08 July : PLR, PFL, and PFL2 Assembly
09 August : Lorem ipsum dolor sit amet
10 September : Hungry Hungry Hipsters

Breakdown

11 July : Lac Promoter and FadD Assembly
12 August : Lorem ipsum dolor sit amet
13 September : Hungry Hungry Hipsters

High-Value Product

14 June : pMBI Assembly
15 July : pMBI and pMevT Assembly
16 August: Troubleshooting Gibson Assembly
17 September: Hungry Hungry Hipsters

Kill Switch

18 July: Hello this is a Description
19 August: Lorem ipsum dolor sit amet
20 September: Hungry Hungry Hipsters

Retrieved from "http://2014.igem.org/Team:CSU_Fort_Collins"