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Oysters & the
Chesapeake

 
Oyster Research and Restoration in U.S. Coastal Waters: Strategies for the Future
September 8-9, 2003 - Annapolis, Maryland

Abstracts
Workgroup: Frontiers in Disease Research

The Role of Iron and Host-Derived Growth Factors in Regulating Gene Expression of the Oyster Parasite Perkinsus marinus: The Basis for Parasite Proliferation Inhibition Strategies

Principal Investigator(s):
Gerardo R. Vasta, Center of Marine Biotechnology, Baltimore, MD, vasta@umbi.umd.edu

Funding Period: 1998-00

  • Recombinant expression and purification of PmSOD1 and PmSOD2 in E. coli. Proof that both enzymes genes encode iron-cofactored SODs (FeSODs).
  • Demonstration that expression of PmSOD1 in yeast causes an FeSOD to be targeted to the mitochondria. This novel finding highlights the potential for PmSOD1 to be a target for intervention.
  • This 'proof of principle' of functional heterologous expression of P. marinus genes shows that a library-based approach, to discover new antioxidant genes, is feasible.
  • Establishment of conditions that allow the growth of P. marinus as microcolonies. This will enable design of mutagenesis screens to identify P. marinus mutants deficient in metabolic or antioxidative pathways.

IMPACTS and/or BENEFITS:

The crystallization and structure determination of recombinant proteins is a prerequisite for developing structure-based inhibition strategies.

Recombinant expression of SODs ultimately led to (1) the crystallization of pure enzymes, and (2) generation of specific antisera against both SODs, and their use in immunofluorescence to discover:
  • The novel localization of the FeSOD to a subcellular compartment suggests that the parasite mitochondrion may itself represent a target for intervention.
Creation and testing of specific mutants in antioxidant pathways is crucial to understanding how this potential virulence-related pathway can be used as a target for intervention.

PROJECT PUBLICATIONS:

Gauthier, J. D. & Vasta, G. R. 2002. Effects of plasma from bivalve mollusk species on the in vitro proliferation of the protistan parasite Perkinsus marinus. Journal of Experimental Zoology 292, 221-30.

Wright, A., Ahmed, H., Gauthier, J., Silva, A. & Vasta, G. 2002. cDNA cloning and characterization of two iron superoxide dismutases from the oyster pathogen Perkinsus marinus. Moleculat and Biochemical Parasitology. 123,73-7

Gauthier, J. D. & Vasta, G. R. Effects of iron on expression of antioxidant genes of the protistan parasite Perkinsus marinus. In preparation.

UM-SG-TS-2003-01 www.mdsg.umd.edu
   
This publication was supported by funds from
the NOAA National Sea Grant College Program and the
Maryland and Virginia Sea Grant College Programs
[Maryland Sea Grant]
 [NOAA]
 [Virginia Sea Grant]

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