Enchytraeus albidus Microarray: Enrichment, Design, Annotation and Database EnchyBASEReportar como inadecuado

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Enchytraeus albidus Oligochaeta is an ecologically relevant species used as standard test organisms for risk assessment. Effects of stressors in this species are commonly determined at the population level using reproduction and survival as endpoints. The assessment of transcriptomic responses can be very useful e.g. to understand underlying mechanisms of toxicity with gene expression fingerprinting. In the present paper the following is being addressed: 1 development of suppressive subtractive hybridization SSH libraries enriched for differentially expressed genes after metal and pesticide exposures; 2 sequencing and characterization of all generated cDNA inserts; 3 development of a publicly available genomic database on E. albidus. A total of 2100 Expressed Sequence Tags ESTs were isolated, sequenced and assembled into 1124 clusters 947 singletons and 177 contigs. From these sequences, 41% matched known proteins in GenBank BLASTX, e-value≤10-5 and 37% had at least one Gene Ontology GO term assigned. In total, 5.5% of the sequences were assigned to a metabolic pathway, based on KEGG. With this new sequencing information, an Agilent custom oligonucleotide microarray was designed, representing a potential tool for transcriptomic studies. EnchyBASE http:-bioinformatics.ua.pt-enchybase- was developed as a web freely available database containing genomic information on E. albidus and will be further extended in the near future for other enchytraeid species. The database so far includes all ESTs generated for E. albidus from three cDNA libraries. This information can be downloaded and applied in functional genomics and transcription studies.

Autor: Sara C. Novais , Joel Arrais, Pedro Lopes, Tine Vandenbrouck, Wim De Coen, Dick Roelofs, Amadeu M. V. M. Soares, Mónica J. B. Am

Fuente: http://plos.srce.hr/


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