Team:UMaryland

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.

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• 2013 SDU Denmark
• 2013 SYSU China
• 2013 Shenxhen BGIG ATCG
• 2013 Colombia Unianades
• 2013 Lethbridge

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