Difference between revisions of "Team:Peshawar"

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    <link rel="icon" type="image/png" href="https://static.igem.org/mediawiki/2016/f/f5/T-Peshawar--favicon-v1-compressor.png">
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<img src="https://static.igem.org/mediawiki/2016/thumb/d/d1/T--Peshawar--iGemers-tint-compressed.png/800px-T--Peshawar--iGemers-tint-compressed.png">
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    <title>iGEM Peshawar 2016</title>
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<a class="blog-ghost-button" href="http://www.synbiokp.com/blog/"  target="_blank">Visit our blog</a>
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<link rel="stylesheet" type="text/css" href="https://2016.igem.org/Template:Peshawar/CSS?action=raw&ctype=text/css" />
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        <ul class="navigation-list">
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            <li><a href="https://2016.igem.org/Team:Peshawar">Home</a></li>
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            <hr>
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            <li><a href="https://2016.igem.org/Team:Peshawar/Description">Project</a></li>
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            <li><a href="https://2016.igem.org/Team:Peshawar/Model">Modeling</a></li>
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            <li><a href="https://2016.igem.org/Team:Peshawar/Notebook">Notebook</a></li>
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            <li><a href="https://2016.igem.org/Team:Peshawar/Safety">Safety</a></li>
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            <li><a href="https://2016.igem.org/Team:Peshawar/Parts">Parts</a></li>
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            <li><a href="https://2016.igem.org/Team:Peshawar/Basic_Part">Basic Part</a></li>
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            <li><a href="https://2016.igem.org/Team:Peshawar/Composite_Part">Composite Part</a></li>
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            <hr>
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            <li><a href="https://2016.igem.org/Team:Peshawar/Team">Team</a></li>
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            <li><a href="https://2016.igem.org/Team:Peshawar/Attributions">Attributions</a></li>
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            <li><a href="https://2016.igem.org/Team:Peshawar/Collaborations">Collaborations</a></li>
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            <hr>
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            <li><a href="https://2016.igem.org/Team:Peshawar/Human_Practices">Human Practices</a></li>
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            <li>
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                <a href="https://2016.igem.org/Team:Peshawar/HP/Silver">Silver</a>
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                <span>  |  </span>
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                <a href="https://2016.igem.org/Team:Peshawar/HP/Gold">Gold</a>
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            </li>
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            <li><a href="https://2016.igem.org/Team:Peshawar/Integrated_Practices">Integrated Practices</a></li>
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            <hr>
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            <li><a href="https://2016.igem.org/Team:Peshawar/Engagement">Public Engagement</a></li>
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            <li><a href="https://2016.igem.org/Team:Peshawar/Awards">Awards</a></li>
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            <hr>
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            <li><a href="https://2016.igem.org/Team:Peshawar/Wiki">How to design an iGEM Wiki</a></li>
  
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                T--Peshawar--blog-round-v2
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            <a href="https://2016.igem.org/Team:Peshawar">
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                <img id="id-logo" class="logo" src="https://static.igem.org/mediawiki/2016/7/7b/T--Peshawar--iGEM-Peshawar-Logo---New-Pakol-v2-2.png">
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<h1> A Biosensor for detecting levels of CO and NOx in vehicle emissions </h1>
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Air pollution is a major global problem, a significant amount of which is caused by vehicular emissions. This problem is particularly prevalent in developing nations, where regulations regarding vehicle emissions are either not present or if present, not properly implemented. Carbon monoxide and oxides of nitrogen are two dangerous constituents of exhaust fumes. Our aim is to produce a portable, quick and easy to use Biosensor device for environmental law enforcement agencies and even consumers that can detect for levels of carbon monoxide and oxides of nitrogen in vehicle exhausts and express corresponding chromoproteins as a result. The system is based on two separate gas-sensing mechanisms that work together to give a range of results. The mechanism sensing for carbon monoxide uses the CooA transcription factor and two CooA dependent promoters: CooF and CooM. The NOx sensing mechanism uses the NsrR repressed promoters, YeaR and NirK, as well as the NorV promoter.</p>
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<h2> Scope of the problem </h2>
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The continuous deterioration of air quality, especially in urban areas, is a matter of global concern. It is responsible for respiratory illnesses which severely affect the quality of life. Air pollution causes an estimated 5.5 million premature deaths every year, worldwide . The number of deaths caused by air pollution is greater than ones caused by other risk factors such as malnutrition, obesity, alcohol and drug abuse. More than 80% of the global urban population is exposed to air that does not meet the WHO air quality standards. The situation is worse for developing countries where air quality levels of 98% of cities exceed WHO limits . </p>
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<p>A World Bank report released in 2014 states that the consequences of air pollution suffered by Pakistan are the highest in South Asia - air pollution is responsible for more deaths than other high profile causes of mortality . With more than 10.6 million vehicles on the road, Karachi, Peshawar and Rawalpindi are among the 20 most polluted cities in the world. According to WHO statistics cited by the World Economic Forum, Karachi is at number 5. The total number of motor vehicles in Karachi is a staggering 1.82 million; with 16,562 new ones hitting the road every month. Although laws and regulations regarding vehicle emissions do exist, the World Bank’s Cleaning Pakistan’s Air report notes implementation as a major problem. Environmental regulatory authorities fail to regulate and keep vehicle emissions in check mainly because of weak implementation and enforcement mechanisms. Having identified this gap, we set on to develop a device that can facilitate the implementation and make measurement of pollutants easier and more accessible.</p>
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<h3>Pollutants</h3>
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Complete combustion, in vehicle engines, give off Carbon dioxide and water as products:
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Fuel (hydrocarbons) + Air (oxygen and nitrogen) --> CO2 + H2O + Unaffected Nitrogen
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However, typical engines are not very efficient and produce Carbon Monoxide and Oxides of Nitrogen along with other pollutants.
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Fuel (hydrocarbons) + Air (oxygen and nitrogen) -->  CO2 + H2O + Unburned Hydrocarbons + <strong>CO (Carbon Monoxide) + NOx (Nitrogen Oxides) </strong>
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<h3>Carbon Monoxide</h3>
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<p>Carbon Monoxide is a partially oxidized product of combustion reaction. It is formed when Hydrocarbon fuel is not completely oxidized to Carbon dioxide. Carbon Monoxide is an odorless, colorless, tasteless and highly poisonous gas. It competes with oxygen and combines with hemoglobin to form carboxyhemoglobin which renders it ineffective for oxygen delivery to body tissues.</p>
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<h3>Oxides of Nitrogen</h3>
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<p>Nitrogen and Oxygen react under high temperature and pressure condition in engine to form several oxides of nitrogen collectively termed as NOX. They are precursors in the formation of ground level ozone which can exacerbate symptoms in respiratory patients. Oxides of nitrogen cause airway irritation and weaken the body's defenses against respiratory infections, such as pneumonia and influenza. They have been linked to the deaths of 23500 patients in UK each year.</p>
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<h2>Current methods for measurement</h2>
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<p>The methods used for measuring gaseous concentration are Non-Dispersive Infrared (NDIR) and Gas Chromatography (GC) for carbon monoxide, and Gas Phase Chemiluminescence for Nitrogen Oxides. All these methods are complex, expensive, and not portable. Our biosensor, on the other hand, will be cheap, easy to use, and highly portable</p>
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<h2>Our Project</h2>
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<p>The whole project centers around a biosensor with two detection systems, one for the CO detection and other for the detection of oxides of nitrogen, particularly, NO and NO2. The detection system for oxides of nitrogen consists of a repressor NsrR, which can be repressed by the inducers NO and NO2, two promoters PYeaR and PNirK, which senses the presence of nitric oxides and expresses GFP and a double terminator. When the oxides of nitrogen is present in the system, it will bind to the NsrR repressor, leading to the activation of promoters PYeaR or PNirK depending on the concentration of nitrogen oxides, that will start expressing GFP, in order to indicate that the oxides of nitrogen are present in the system.</p>
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<p>The second system that works for the detection of CO, consists of an activator CooA, which is normally present in an inactive form in the system and requires the presence of two molecules of CO in order to get activated. The system also comprises of two promoters including pCooF and pCooM that is sensitive to the presence of CO, pCooF is a strong promoter and requires the attachment of two molecules of CO, while pCooM is a weak promoter where the attachment of eight molecules of CO is necessary in order to express the GFP. The promoters are followed by the double terminator. When CO is present in the system, two molecules of CO will bind to the inactive activator, thus making it active, the activated CO + CooA complex will now bind to the promoters, the activated promoter will lead to the production of GFP, that has the tendency to indicate that CO is present in the system.</p>
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<p>The system is composed of a collaboration of six different devices in an E. coli K12 Strain. The devices includes:</p>
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                            <h2 id="heading-line-1"><span id="inspector">Inspector</span> <span id="NO">NO</span>.<span id="CO">CO</span>.<span id="li">li</span> - A biosensor for detection</h2>  
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<li><strong>CooA device</strong>
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                            <h2 id="heading-line-2">of levels of CO and NOx in vehicle emissions</h2>
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<p>The CooA device comprises of four parts and is responsible for the production of CooA activator protein. The parts are:</p>
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<li>a constitutive promoter</li>
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<li>RBS</li>
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<li>a coding region for CooA protein</li>
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<li>a double terminator</li>
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<li><strong>pCooF device</strong>
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<p>The pCooF device codes for the strong promoter which senses the presence of CO, and then expresses GFP. It is composed of four parts:</p>
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<li>a pCooF promoter</li>
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<li>RBS</li>
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<li>coding region for desired GFP</li>
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<li>a double terminator</li>
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<li><strong>pCooM device</strong>
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<p>The pCooM device produces pCooM, which is a weak promoter, with a GFP expression. It uses four parts:</p>
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<li>pCooM promoter</li>
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<li>RBS</li>
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<li>coding region for desired GFP</li>
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<li>a double terminator</li>
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<li><strong>NsrR device</strong>
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<p>NsrR device is composed of four parts and is responsible for the production of NsrR repressor protein. The parts includes:</p>
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<li>a constitutive promoter</li>
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<li>RBS</li>
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<li>coding region for NsrR protein</li>
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<li>a double terminator</li>
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<li><strong>PYeaR device</strong>
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<p>The PYeaR device code for PYeaR promoter, which when interacts with oxides of nitrogen produces GFP. It is composed of four parts:</p>
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<ul>
+
−
<li>PYeaR promoter</li>
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<li>RBS</li>
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<li>coding region for GFP</li>
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<li>a double terminator</li>
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</li>
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<li><strong>PNirK device</strong>
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<p>The PNirK device is used for the GFP expression. It is composed of four parts:</p>
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<li>PNirK promoter</li>
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<li>RBS</li>
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<li>coding region for GFP</li>
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<li>a double terminator</li>
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</ul>
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</li>
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<o/l>
+
  
  
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Revision as of 14:55, 19 October 2016

<!DOCTYPE html> iGEM Peshawar 2016

Inspector NO.CO.li - A biosensor for detection

of levels of CO and NOx in vehicle emissions





Icon credits: Luis Prado, Martin Vanco, Vaibhav Radhakrishnan, Alex Tai, Anton, Alex Kwa, Nick Abrams



Reach Us

Institute of Integrative Biosciences,
CECOS University of IT & Emerging Sciences,
Phase 6, F-5, Hayatabad, Peshawar, Pakistan

iib@cecos.edu.pk

+92 91 5860291

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