Team:Leeds

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Who the heck are Leeds?

We are the Leeds 2013 iGEM team, We are a group of undergrads from various courses all with a passion for synthetic biology!

So what's the Big Idea?

Like the hunting dog, the Beagle, our Micro-beagle is a biosensor designed to hunt down pathogens. Micro-beagle utilises the Cpx pathway and GFP, which, when it fluoresces can be seen with the naked eye. In order to achieve this we will have designed our biosensor to physically bind to the antigen of interest. This will induce membrane stress which activates the Cpx pathway. By putting the GFP gene downstream of the Cpx promoter, GFP protein will be made when membrane stress occurs and consequently the cells will fluoresce when the pathogen is present in the solution.
As a proof of concept, first we will be using silica beads as a pathogen analogue. In order to bind to the beads will be express a silica binding peptide on the surface of the cells using ice nucleation protein. INP, a BioBrick we decided to use from the registry, expresses any gene sequence placed at its C terminus on the cell surface. The theory is that the silica binding peptide sequence can be easily swapped for a sequence of any antigen binding moiety and therefore enable us to detect any pathogen, simply, quickly and cheaply.

Geneious, our fine sponsors and suppliers of software

Bioline, our fine sponsors and suppliers of equipment

Qiagen, our fine sponsors and suppliers of PCR kits

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 ==So what's the [[Team:Leeds/Project | Big Idea]]?==
-Our initial idea is to design a system capable of testing water for the presence of pathogenic bacteria, this system will initially be modeled using silica beads and a Si4 attachment peptide. Our idea is to use Ice Nucleation protein to present the Si4 peptide on the outer membrane of our ''E. coli'' in conjunction with the membrane stress modulator, CpxR promoter, linked to Green Fluorescent protein. The binding of ''E.coli'' to the silica beads will hopefully cause enough membrane stress to induce the CpxR promoter and therefore production of GFP. In addition to this we will be incorporating the LuxI quorum sensing pathway to another GFP tag resulting in massive signal amplification between all of our bacteria in solution creating an all of nothing response to the signal.
+Like the hunting dog, the Beagle, our Micro-beagle is a biosensor designed to hunt down pathogens. Micro-beagle utilises the Cpx pathway and GFP, which, when it fluoresces can be seen with the naked eye.
+In order to achieve this we will have designed our biosensor to physically bind to the antigen of interest. This will induce membrane stress which activates the Cpx pathway. By putting the GFP gene downstream of the Cpx promoter, GFP protein will be made when membrane stress occurs and consequently the cells will fluoresce when the pathogen is present in the solution.
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-If this is system is successful we can use directed evolution of our peptide tag, presented by INP, to have it attach to pathogenic bacteria resulting in the same GFP response.
+As a proof of concept, first we will be using silica beads as a pathogen analogue. In order to bind to the beads will be express a silica binding peptide on the surface of the cells using ice nucleation protein. INP, a BioBrick we decided to use from the registry, expresses any gene sequence placed at its C terminus on the cell surface. The theory is that the silica binding peptide sequence can be easily swapped for a sequence of any antigen binding moiety and therefore enable us to detect any pathogen, simply, quickly and cheaply.
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