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Electronic Journal of Biotechnology
Universidad Católica de Valparaíso
ISSN: 0717-3458
Vol. 14, No. 1, 2011
Bioline Code: ej11008
Full paper language: English
Document type: Short Communication
Document available free of charge

Electronic Journal of Biotechnology, Vol. 14, No. 1, 2011

 en Suppression subtractive hybridization PCR isolation of cDNAs from a Caribbean soft coral
Lopez, Jose V.; Ledger, Angela; Santiago-Vázquez, Lory Z.; Pop, Mihai; Sommer, Dan D.; Ranzer, Llanie K.; Feldman, Robert A. & Kerr, Russell G.

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 check for this species in other resources and the sea fan Gorgonia ventalina check for this species in other resources 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, GH611838, and HO061755-HO062154]. 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.

Keywords
cDNA, Erythropodium caribaeorum, EST, Gorgonia ventalina, gorgonian, sea fan.

 
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