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Article
Suppression Subtractive Hybridization PCR Isolation of cDNAs from a Caribbean Soft Coral
Electronic Journal of Biotechnology
  • Jose V. Lopez, Nova Southeastern University
  • Angela Ledger, Florida Atlantic University
  • Lory Z. Santiago-Vázquez, Florida Atlantic University
  • Mihai Pop, University of Maryland
  • Dan D. Sommer, University of Maryland
  • Llanie K. Ranzer, Florida Atlantic University
  • Robert A. Feldman, SymBio Corporation
  • Russell G. Kerr, Florida Atlantic University
ORCID
0000-0002-1637-4125
ResearcherID
F-8809-2011
Document Type
Article
Publication Date
1-15-2011
Keywords
  • cDNA,
  • Erythropodium caribaeorum,
  • EST,
  • Gorgonia ventalina,
  • Gorgonian,
  • Sea fan
Abstract

Transcriptomic studies of marine organisms are still in their infancy. A partial, subtracted expressed sequence tag (EST) library of the Caribbean octocoral Erythropodium caribaeorum and the sea fan Gorgonia ventalina has been analyzed in order to find novel genes or differences in gene expression related to potential secondary metabolite production or symbioses. This approach entails enrichment for potential non-"housekeeping" genes using the suppression subtractive hybridization (SSH) polymerase chain reaction (PCR) method. More than 500 expressed sequence tags (ESTs) were generated after cloning SSH products, which yielded at least 53 orthologous groups of proteins (COGs) and Pfam clusters, including transcription factors (Drosophila Big Brother), catalases, reverse transcriptases, ferritins and various "hypothetical" protein sequences. A total of 591 EST sequences were deposited into GenBank [dbEST: FL512138 - FL512331, and GH611838]. The results represent proof of concept for enrichment of unique transcripts over housekeeping genes, such as actin or ribosomal genes, which comprised approximately 17% of the total dataset. Due to the gene and sequence diversity of some ESTs, such sequences can find utility as molecular markers in current and future studies of this species and other soft coral biogeography, chemical ecology, phylogenetics, and evolution.

DOI
10.2225/vol14-issue1-fulltext-3
Comments

©2011 by Pontificia Universidad Catolica de Valparaiso - Chile

Additional Comments
Florida Sea Grant grant #s: R/LR-MB-23, R/LR-MB-14; NSF grant #s: 0310283, 0514500; NIH grant #: R01-HG-0004885
Citation Information
Lopez, Jose V., Angela Ledger, Lory Z. Santiago-Vazquez, Mihai Pop, Dan D. Sommer, Llanie K. Ranzer, Robert A. Feldman, and Russell G. Kerr. "Suppression Subtractive Hybridization PCR Isolation of cDNAs from a Caribbean Soft Coral." Electronic Journal of Biotechnology. no. 1 (2011). 10.2225/vol14-issue1-fulltext-3.