Team:UMaryland

From 2014.igem.org

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Revision as of 00:53, 15 August 2014


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Project Description

The Chesapeake Bay is one of the most treasured wildlife biomes in Maryland. However, overfishing, pollution, and the need for commercial trade has disrupted this habitat.

A major biological reason for this decline in the Eastern oyster population has been an oyster pathogen called Perkinsus marinus, a unicellular protist which causes an infection known as Dermo in oyster tissues. P. marinus has been shown to infest oysters by entering their hemocytes via induced phagocytosis. When inside the hemocyte, this parasite is capable of resisting the reactive oxygen species generated by the oyster, allowing it to proliferate inside the oyster cells. The growth of P. marinus is eventually fatal to the oyster.

It is believed that P. marinus enters oyster hemocytes utilizing a galectin called CvGal1 that expressed and secreted by hemocytes. Our goal is to exploit the ability of CvGal1 to bind to P. marinus in order to design a biosensor for the detection of this pathogen. As CvGal1 is a soluble protein, we need a mechanism to anchor it to the outer cell membrane. To accomplish this, we are interested in fusion CvGal1 to the C-terminus of the transmembrane domain of OmpA, an integral outer membrane protein. By generating an OmpA-CvGal1 fusion protein, we hope to express this protein on the E. coli surface in order to trigger a signal transduction cascade within the bacterium via the Cpx two component system. This signal cascade would lead to the production of a signal molecule, such as GFP. Thus, P. marinus binding to CvGal1 would trigger the creation of a visual marker for the parasite.

We are currently also investigating the potential of generating a response from E. coli to mitigate the pathogenicity of P. marinus. This could potentially be done by generating cleavage enzymes to digest ligands on the surface of P. marinus that bind to CvGal1, reducing the ability of oysters to recognize and phagocytose the pathogen.

Although laboratory methods for Dermo detection using ELISA exist, they are currently not capable of detecting the disease in real time. In the time needed to conduct a laboratory test, Dermo could have easily spread to new locations. This biosensor would help to accurately pinpoint Dermo infestation levels in real time. If we can pinpoint the location of P. marinus, then we can ultimately develop a biosensor that can react to Dermo with targeted destruction.

Requirements

There are a few wiki requirements teams must follow:

  • All pages, images and files must be hosted on the 2014.igem.org server.
  • All pages must be created under the team’s name space.
  • As part of your documentation, keep the links from the menu to the left.
  • Do not use flash in wiki code.
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Visit the Wiki How To page for a complete list of requirements, tips and other useful information.

Tips

We are currently working on providing teams with some easy to use design templates.
In the meantime you can also view other team wikis for inspiration! Here are some very good examples

For a full wiki list, you can visit iGEM 2013 web sites and iGEM 2012 web sites lists.

  • State your accomplishments! Tell people what you have achieved from the start.
  • Be clear about what you are doing and what you plan to do.
  • 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 2013 calendar
  • Have lots of fun!