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Analysis of Expressed Sequence Tags from the Harmful Alga, Prymnesium parvum (Prymnesiophyceae, Haptophyta)

Abstract

A cDNA library was constructed from late log-phase cultures of the ichthyotoxin-producing haptophyte, Prymnesium parvum. Approximately 6,300 single-pass 5′ nucleotide sequences (out of nearly 8,000 randomly isolated clones) achieved length and quality criteria for incorporation into an expressed sequence tag (EST) database. Collectively, 3,415 unique genes (tentative unigenes [TUGs]) were assembled. Database searches revealed approximately 1,100 TUG (34%) that encode proteins of known or putative function (expect values E ≤ 1.0 × 10−10), representing most functional classes of proteins and many metabolic pathways. In addition to proteins involved in photosynthesis and protein synthesis/degradation, one of the most frequently encountered transcripts putatively encodes an ABC-type phosphate transport system component. Active (and efficient?) phosphate transport may correlate with rapid growth, perhaps explaining how this alga out-competes other species when it blooms and becomes toxic under low-phosphate conditions. The majority of TUGs, including 12 of the 50 most commonly encountered transcripts, encode potentially novel proteins, some of which are probably involved in synthesis and secretion of the unique prymnesin toxins. This work suggests that future studies on the expression of many of these genes during rapid growth should provide insight into the genetic basis of bloom formation and toxicity.

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Acknowledgments

The author is particularly grateful to colleague and collaborator Prof. David. L. Herrin for innumerable suggestions, ideas, and discussions; to David Buzan, T.O. Smith, and especially Dr. Loraine Fries (Texas Parks and Wildlife Department) for support and for generously making available their resources and talents; to Lindsay Bissonnet and Schonna Manning for outstanding technical assistance; to Cecil Harkey and Alison Mangum (U.T. DNA Facility) for unflagging cooperation with the sequencing; to Dr. Jim Leebens-Mack (Pennsylvania State University) for many valuable suggestions and help managing and analyzing the sequence data; and to Prof. Lukas Mueller and Teri Solow (Plant Genome Network, Cornell University) for availing the use of their pipeline and for generous help with creating and maintaining the EST database. This work was supported by the U.S. Fish and Wildlife Service State Wildlife Grant T-14-P and the Texas Parks and Wildlife Department under grant no. 128312.

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Correspondence to John W. La Claire II.

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La Claire, J.W. Analysis of Expressed Sequence Tags from the Harmful Alga, Prymnesium parvum (Prymnesiophyceae, Haptophyta). Mar Biotechnol 8, 534–546 (2006). https://doi.org/10.1007/s10126-005-5182-2

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Keywords

  • expressed sequence tag
  • functional genomics
  • Haptophyta
  • metabolism
  • Prymnesium parvum