Suppression subtractive hybridization PCR isolation of cDNAs from a Caribbean soft coral
Electron. j. biotechnol
; 14(1): 8-9, Jan. 2011. ilus, tab
Article
in En
| LILACS
| ID: lil-591926
Responsible library:
CL1.1
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, 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 percent 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.
Key words
Full text:
1
Collection:
01-internacional
Database:
LILACS
Main subject:
DNA, Complementary
/
Anthozoa
Limits:
Animals
Language:
En
Journal:
Electron. j. biotechnol
Journal subject:
BIOTECNOLOGIA
Year:
2011
Document type:
Article
/
Project document
Affiliation country:
United States
Country of publication:
Chile