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The main objective of this study is to generate expressed sequence tags (EST) data from single-pass sequencing of clones from P. marinus exposed to conditions that have been demonstrated to increase parasite virulence, such as high temperature, salinity, and iron levels. We will construct normalized P. marinus cDNA libraries using cultures grown under diverse physical factors (high temperature, salinity, iron) and biological factors (presence of oyster serum). EST sequences will be generated using two approaches (i) clones directly sequenced from the cDNA libraries, and (ii) clones sequenced from subtracted cDNA libraries using P. marinus grown in standard culture medium. The characterization of the expressed genome (ESTs) may result in the identification of suitable targets for genetic intervention. Key gene products for Perkinsus fitness, survival, and/or interaction with C. virginica should be susceptible to being disabled by genetically engineered and/or selected oysters that carry the appropriate genotype, or by the identification of metabolic pathways in Perkinsus that may constitute targets for chemotherapy.
Pecher, WT; Alavi, MR; Schott, EJ; Fernandez-Robledo, JA; Roth, L; Berg, ST; Vasta, GR. 2008. Assessment of the northern distribution range of selected Perkinsus species in eastern oysters (Crassostrea virginica) and hard clams (Mercenaria mercenaria) with the use of PCR-based detection assays. Journal of Parasitology94(2):410 -422. doi:10.1645/GE-1282.1. UM-SG-RS-2008-12.