Development of reference transcriptome de novo construction database of Brown-marbled grouper (Epinephelus fuscoguttatus) spleen in response to vibriosis infection. Epinephelus fuscoguttatus

We constructed a reference transcriptome for the Brown-marbled Grouper E. fuscoguttatus, representing a model of gene expression in infection mode. The infection were from vibriosis pathogens infected the fish at fingerlings stage, this transcriptome was built with added intention of serving as community resources to facilitates investigations into the molecular basis of innate immunity related gene expression. Assembly of Illumina sequenced reads from normalized library resulted in 20,795 contigs, with putative homologues identified from Master Salmo Salar transcriptome assembly deposited in Genebank. To test the utility of this reference transcriptome for microarray experiments, 15K probes designed on microarray slides and hybridized with RNA of challenge and non-challenge fish at selected times. This assay allowed us characterized gene expression during infection stage, tabulated immune-related genes and predicted immune response pathways. Further, confirmation of immune related genes expression was done using qualitative real-time PCR and the interest genes are set of molecular markers. This molecular marker was then tested in another in-vivo assay, Brown-marbled grouper larvae challenge with Vibrio harveyi and shown to be good indicator of immune response.