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Revision as of 15:22, 29 June 2017
Project Description
Our project idea is to develop a biosensor to detect the presence of the bacteria Campylobacter. 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 Pseudomonas fluorescens and expressing this in our chassis organism, Escherichia Coli, 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 campylobacter 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.
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 campylobacter.
To provide increased specificity to our biosensor, we are aiming to detect autoinducer-2, a quorum sensing molecule released by campylobacter. 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.
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.
Welcome to iGEM 2017!
Your team has been approved and you are ready to start the iGEM season!
Before you start:
Please read the following pages:
Styling your wiki
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.
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.
Wiki template information
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 Pages for awards 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!
Editing your wiki
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!
Use WikiTools - Edit in the black menu bar to edit this page
Tips
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:
- State your accomplishments! Tell people what you have achieved from the start.
- Be clear about what you are doing and how you plan to do this.
- You have a global audience! Consider the different backgrounds that your users come from.
- Make sure information is easy to find; nothing should be more than 3 clicks away.
- Avoid using very small fonts and low contrast colors; information should be easy to read.
- 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 iGEM 2017 calendar
- Have lots of fun!
Inspiration
You can also view other team wikis for inspiration! Here are some examples:
Uploading pictures and files
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.
When you upload, set the "Destination Filename" to T--YourOfficialTeamName--NameOfFile.jpg. (If you don't do this, someone else might upload a different file with the same "Destination Filename", and your file would be erased!)
UPLOAD FILES
