Difference between revisions of "Team:Glasgow"

 
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<i>Campy</i>LOCATOR
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Detection of <i>Campylobacter jejuni</i> for the prevention of food poisoning </i>
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<h1> Project Description </h1>
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<p> Our project idea is to develop a biosensor to detect the presence of the bacteria <i>Campylobacter</i>. This sensor will utilise the rare sugar xylulose, which is found in the polysaccharide capsule of campylobacter and is released when the bacteria is run through an acidic solution. By exploiting the mannitol operon that is present in the bacteria <i>Pseudomonas fluorescens</i> and expressing this in our chassis organism, <i>Escherichia Coli</i>, we will produce a biosensor that will express the reporter molecule Green Fluorescent Protein (GFP) when xylulose interacts with the repressor molecule of the mannitol operon. Additional sub-projects will include; investigating the quorum sensing mechanisms in <i>campylobacter</i> to increase the specificty of our sensor, developing hardware to produce a functioning biosensor and investigating the legal and ethical issues associated with our project.</p>
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<p>The mannitol operon contains a promoter, Pe, which is regulated by xylulose, sorbitol and mannitol. The operon typically regulates the expression of genes required for mannitol consumption. By hijacking this xylulose regulated promoter we hope to develop a construct capable of activating GFP in the presence of xylulose. A schematic of the construct is shown in the diagram opposite. We are also looking at the Arabinose Operon and how it interacts with xylulose as a potential alternative if we cannot get our original construct to work. We will perform saturation mutagenesis on the sugar binding site of the AraC protein with the hope of finding a mutant which will interact with the pBAD promoter to cause initiation of gene expression when xylulose is present which would lead to the detection of <i>campylobacter</i>.</p>
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<i>Campylobacter </i> species, especially <i>Campylobacter jejuni</i>, are bacterial pathogens responsible for the majority of food poisoning in the UK. This bacteria is considered to cause over 280,000 cases of bacterial gastroenteritis annually. Although also found on red meat, unpasteurized milk and unfiltered water, 4 out of 5 cases of campylobacteriosis come from contaminated chickens. During the slaughter process the bacterium gets transferred onto the chicken skin and is often found on fresh retail chickens and their packaging. The most important transmission route is consumption of undercooked chicken or other foods that get cross-contaminated from raw poultry meat. </p>
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<p class="copy"> Current detection systems for the pathogen are time-consuming, expensive and inaccessible for everyday users. We therefore aimed to develop a new, cheaper and faster system for detection of <i>C. jejuni</i> using synthetic biology. We approached this by genetically engineering <i>Escherichia coli</i> to serve a function of a dual-input biosensor. The bacterium will sense bacterial quorum sensing signals and xylulose - a sugar naturally present in the outer capsule of <i>Campylobacter jejuni</i>, to detect the pathogen on a swabbed surface. </p>
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<a href="https://2017.igem.org/Team:Glasgow/Accomplishments">Our Accomplishments at the Jamboree!</a>
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<p> To provide increased specificity to our biosensor, we are aiming to detect autoinducer-2, a quorum sensing molecule released by <i>campylobacter</i>. We aim to insert the LsrA promotor followed by YFP into a plasmid which turns on when autoinducer-2 is present. For our proof of concept, we need to work with xylulose. However, xylulose is rare and, as such, is expensive. Therefore, we will synthesise xylulose by utilising a metabolic step in the bacterial pentose pathway. The enzyme Xylose Isomerase can be purified from E. coli, and used to convert the inexpensive sugar xylose in to xylulose. </p>
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<p>For the engineering part of the project, we are aiming to build a functional biosensor that will be able to prove our construct. We will use technologies such as microfluidics and ultrafiltration to produce a sensor that will use our modified E.coli to give a visual indication of the presence of campylobacter. In terms of human practices, we will be adopting a double approach; one that focuses on the legal and political issues concerning EU legislation, which will affect our ability to manufacture a biosensor outside a properly licensed laboratory; and one on ethical issues concerning the impact that food safety has on human health. This second approach will mainly aim to raise awareness among the public on preventative measures in order to avoid campylobacter poisoning.</p>  
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<a href="https://2017.igem.org/Team:Glasgow/mtlR">Engineering <i>E. coli</i> to Respond to <i>Campylobacter</i>-Associated Sugar</a>
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<h1> Welcome to iGEM 2017! </h1>
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<p>Your team has been approved and you are ready to start the iGEM season! </p>
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<a href="https://2017.igem.org/Team:Glasgow/araC">Expanding the Biosensor parts Toolkit, using <i>araC</i> Mutants</a>
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<h5>Before you start: </h5>
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<p> Please read the following pages:</p>
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<li>  <a href="https://2017.igem.org/Competition">Competition Hub</a> </li>
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<li> <a href="https://2017.igem.org/Competition/Deliverables/Wiki">Wiki Requirements page</a></li>
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<li> <a href="https://2017.igem.org/Resources/Template_Documentation">Template documentation</a></li>
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<h5> Styling your wiki </h5>
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<a href="https://2017.igem.org/Team:Glasgow/Hardware">Designing a Functional Prototype Biosensor device</a>  
<p>You may style this page as you like or you can simply leave the style as it is. You can easily keep the styling and edit the content of these default wiki pages with your project information and completely fulfill the requirement to document your project.</p>
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<p>While you may not win Best Wiki with this styling, your team is still eligible for all other awards. This default wiki meets the requirements, it improves navigability and ease of use for visitors, and you should not feel it is necessary to style beyond what has been provided.</p>  
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<a href="https://2017.igem.org/Team:Glasgow/Outreach">Engaging with the world about <i>Campylobacter</i></a>
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<h5> Wiki template information </h5>
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<p>We have created these wiki template pages to help you get started and to help you think about how your team will be evaluated. You can find a list of all the pages tied to awards here at the <a href="https://2017.igem.org/Judging/Pages_for_Awards">Pages for awards</a> link. You must edit these pages to be evaluated for medals and awards, but ultimately the design, layout, style and all other elements of your team wiki is up to you!</p>
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<h5> Editing your wiki </h5>
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<p>On this page you can document your project, introduce your team members, document your progress and share your iGEM experience with the rest of the world! </p>
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<p> <a href="https://2017.igem.org/wiki/index.php?title=Team:Example&action=edit"> </a>Use WikiTools - Edit in the black menu bar to edit this page</p>
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<h5>Tips</h5>
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<p>This wiki will be your team’s first interaction with the rest of the world, so here are a few tips to help you get started: </p>
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<li>State your accomplishments! Tell people what you have achieved from the start. </li>
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<li>Be clear about what you are doing and how you plan to do this.</li>
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<li>You have a global audience! Consider the different backgrounds that your users come from.</li>
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<li>Make sure information is easy to find; nothing should be more than 3 clicks away.  </li>
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<li>Avoid using very small fonts and low contrast colors; information should be easy to read.  </li>
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<li>Start documenting your project as early as possible; don’t leave anything to the last minute before the Wiki Freeze. For a complete list of deadlines visit the <a href="https://2017.igem.org/Calendar">iGEM 2017 calendar</a> </li>
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<li>Have lots of fun! </li>
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<h5>Inspiration</h5>
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<p> You can also view other team wikis for inspiration! Here are some examples:</p>
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<li> <a href="https://2014.igem.org/Team:SDU-Denmark/"> 2014 SDU Denmark </a> </li>
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<li> <a href="https://2014.igem.org/Team:Aalto-Helsinki">2014 Aalto-Helsinki</a> </li>
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<li> <a href="https://2014.igem.org/Team:LMU-Munich">2014 LMU-Munich</a> </li>
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<li> <a href="https://2014.igem.org/Team:Michigan"> 2014 Michigan</a></li>
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<li> <a href="https://2014.igem.org/Team:ITESM-Guadalajara">2014 ITESM-Guadalajara </a></li>
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<li> <a href="https://2014.igem.org/Team:SCU-China"> 2014 SCU-China </a></li>
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<h5> Uploading pictures and files </h5>
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<p> You can upload your pictures and files to the iGEM 2017 server. Remember to keep all your pictures and files within your team's namespace or at least include your team's name in the file name. <br />
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When you upload, set the "Destination Filename" to <br><code>T--YourOfficialTeamName--NameOfFile.jpg</code>. (If you don't do this, someone else might upload a different file with the same "Destination Filename", and your file would be erased!)<br><br>
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UPLOAD FILES
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Latest revision as of 21:00, 15 December 2017

Glasgow iGEM 2017
CampyLOCATOR
Detection of Campylobacter jejuni for the prevention of food poisoning
Project Description

Campylobacter species, especially Campylobacter jejuni, are bacterial pathogens responsible for the majority of food poisoning in the UK. This bacteria is considered to cause over 280,000 cases of bacterial gastroenteritis annually. Although also found on red meat, unpasteurized milk and unfiltered water, 4 out of 5 cases of campylobacteriosis come from contaminated chickens. During the slaughter process the bacterium gets transferred onto the chicken skin and is often found on fresh retail chickens and their packaging. The most important transmission route is consumption of undercooked chicken or other foods that get cross-contaminated from raw poultry meat.

Current detection systems for the pathogen are time-consuming, expensive and inaccessible for everyday users. We therefore aimed to develop a new, cheaper and faster system for detection of C. jejuni using synthetic biology. We approached this by genetically engineering Escherichia coli to serve a function of a dual-input biosensor. The bacterium will sense bacterial quorum sensing signals and xylulose - a sugar naturally present in the outer capsule of Campylobacter jejuni, to detect the pathogen on a swabbed surface.

Using synthetic biology to create a biosensor for detection of Campylobacter jejuni, a bacteria linked to food poisoning.