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<h3 id="header">A Brief Description:</h3>
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  <p>Nitrogen is a highly abundant element in the Earth’s atmosphere. Nitrogen is also essential for life. However, atmospheric nitrogen (N2) is not useful in living processes. Reactive nitrogen such as Ammonia or Nitrate are reactive forms of nitrogen used by plants and animals. To maximize crop yield, many farmers add nitrogen based fertilizer to their crops. Oftentimes excess nitrogen in the soil is washed away by rain into other natural water systems. This nitrogen runoff can and does have devastating effects on the biodiversity in waterways. Excess nitrogen in waterways can cause eutrophication, where algae takes advantage of the excess nitrogen and feed on it causing much larger algal blooms. The algal blooms eventually lead to a dramatic reduction of dissolved oxygen in the water, which causes a dead zone where little to no aquatic animals and plants are able to survive. An example of a dead zone largely caused by excess reactive nitrogen is the dead zone in the Gulf of Mexico which measures approximately 5,500-6,500 square miles (Gulf of Mexico Dead Zone).</p>
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<p>The Haber-Bosch process is used to produce reactive nitrogen, specifically for fertilizers. This process produces approximately 100Tg of reactive nitrogen per year (Chen, Ling-Hsiang). It is clear we can effectively produce reactive nitrogen, however, we need to clean up after ourselves by finishing the the nitrogen cycle. There is a lot more reactive nitrogen being introduced to the environment, but the rate of denitrification has essentially stayed the same. Our team will focus on denitrification, in an effort to restore safer nitrogen levels in waterways and bodies of water and ultimately to help make the completion of the nitrogen cycle more efficient.</p>
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<p>How we plan to achieve denitrification:</p>
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<div class="drawing board" style=" background-color: #2e3141; padding-top:50px;">
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                      <center><h2 style="height:100px; padding-top:5px"><font size="+10">Description</font></h2></center>
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                      <center><font color="silver">Our project, "Knockout Nitrate", is a developing project with the aim to use synthetic biology to balance the nitrogen cycle. </font></center>
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<p><center><font color="silver"><h2>Background</h2></font></center></p>
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<p><center><font color="silver">Nitrogen is a highly abundant element in the Earth’s atmosphere. Nitrogen is also essential for life. However, atmospheric nitrogen (N2) is not useful in living processes. Humans have developed processes to convert unreactive atmospheric nitrogen into a usable form of nitrogen such as Ammonia or Nitrate for plants and animals (reactive nitrogen). To maximize crop yield, many farmers add nitrogen based fertilizer to their crops. Oftentimes excess nitrogen in the soil is washed away by rain into other natural water systems. This nitrogen runoff can and does have devastating effects on the biodiversity in waterways.</font></center></p>
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<p><center><font color="silver">To read more background information about the nitrate problem, click the image below.</font></center></p>
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<center><a href="https://2016.igem.org/Team:UNebraska-Lincoln/background"><img src="https://static.igem.org/mediawiki/2016/6/67/T--UNebraska-Lincoln--background.png" style="width:40%; height:auto;" alt="image"/></a></center>
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<br><br>
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<p><center><font color="silver"><h2>Project Design</h2></font></center></p>
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<p><center><font color="silver">In order to combat the nitrate problem, we aimed to design a strain of <i>E. coli</i> capable of reducing these excess nitrate ions. By isolating a gene that codes of periplasmic nitrate reductases, we began the process of designing our synthetic organism.</font></center></p>
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<p><center><font color="silver">To read more information about the design of our system, click the image below.</font></center></p>
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<center><a href="https://2016.igem.org/Team:UNebraska-Lincoln/design"><img src="https://static.igem.org/mediawiki/2016/0/0f/T--UNebraska-Lincoln--design.png" style="width:40%; height:auto;" alt="image"/></a></center>
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<p><center><font color="silver"><h2>Experiments for Our System</h2></font></center></p>
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<p><center><font color="silver">Throughout the summer and into the fall, we implemented our design into the laboratory. We isolated and synthesized our periplasmic nitrate reductase genes, as well as developed a kill switch and safety-assurance cases</font></center></p>
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<p><center><font color="silver">To read more information on the application of our design, click the image below.</font></center></p>
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<center><a href="https://2016.igem.org/Team:UNebraska-Lincoln/experiments"><img src="https://static.igem.org/mediawiki/2016/1/16/T--UNebraska-Lincoln--experiments.png" style="width:40%; height:auto;" alt="image"/></a></center>
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<p><center><font color="silver"><h2>Recharacterization of K381001</h2></font></center></p>
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<p><center><font color="silver">In the design of our project, we implemented a nitrate-sensitive kill switch. This took advantage of the fact that the yeaR promoter, originally submitted by the Edinburgh iGEM Team in 2009 (<a href="http://parts.igem.org/Part:BBa_K216005">K216005</a>) was active only in the presence of nitrate ion. We decided to begin our project by determining sensitivity of this promoter the concentrations of nitrate that would be present in waterways. To do this, we recharacterized the composite part <a href="http://parts.igem.org/Part:BBa_K381001">K381001</a>, a PyeaR-GFP nitrate sensor.  You can read more about our rechracterization of the PyeaR-GFP Biobrick <a href="http://parts.igem.org/Part:BBa_K381001:Experience#UNebraska-Lincoln_2016">Here</a>.</font></center></p>
  
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<p>Denitrification Process: NO3 → NO2 → NO → N2O → N2
 
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Latest revision as of 02:55, 20 October 2016

<!DOCTYPE HTML> Elements - Solid State by HTML5 UP

Description

Our project, "Knockout Nitrate", is a developing project with the aim to use synthetic biology to balance the nitrogen cycle.

Background

Nitrogen is a highly abundant element in the Earth’s atmosphere. Nitrogen is also essential for life. However, atmospheric nitrogen (N2) is not useful in living processes. Humans have developed processes to convert unreactive atmospheric nitrogen into a usable form of nitrogen such as Ammonia or Nitrate for plants and animals (reactive nitrogen). To maximize crop yield, many farmers add nitrogen based fertilizer to their crops. Oftentimes excess nitrogen in the soil is washed away by rain into other natural water systems. This nitrogen runoff can and does have devastating effects on the biodiversity in waterways.

To read more background information about the nitrate problem, click the image below.

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Project Design

In order to combat the nitrate problem, we aimed to design a strain of E. coli capable of reducing these excess nitrate ions. By isolating a gene that codes of periplasmic nitrate reductases, we began the process of designing our synthetic organism.

To read more information about the design of our system, click the image below.

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Experiments for Our System

Throughout the summer and into the fall, we implemented our design into the laboratory. We isolated and synthesized our periplasmic nitrate reductase genes, as well as developed a kill switch and safety-assurance cases

To read more information on the application of our design, click the image below.

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Recharacterization of K381001

In the design of our project, we implemented a nitrate-sensitive kill switch. This took advantage of the fact that the yeaR promoter, originally submitted by the Edinburgh iGEM Team in 2009 (K216005) was active only in the presence of nitrate ion. We decided to begin our project by determining sensitivity of this promoter the concentrations of nitrate that would be present in waterways. To do this, we recharacterized the composite part K381001, a PyeaR-GFP nitrate sensor. You can read more about our rechracterization of the PyeaR-GFP Biobrick Here.