Identification and expression profile of multiple genes in channel catfish fry 10 min after modified live...

Using PCR-select subtractive cDNA hybridization technique, 32 expressed sequence tags (ESTs) were isolated from 96 clones of a channel catfish (Ictalurus punctatus) fry subtractive library 10min post-vaccination with a modified live Flavobacterium columnare vaccine. The transcription levels of the 32 ESTs in response to F. columnare vaccination were then evaluated by quantitative PCR (QPCR). Of the 32 ESTs, 28 were upregulated in at least one vaccinated fish. Of the 28 upregulated ESTs, 12 were consistently induced at least 2-fold higher in vaccinated fish compared to unvaccinated control fish. Of the 12 upregulated genes, three (triglyceride lipase, PIKK family atypical protein kinase, and CCR4-NOT transcription complex subunit 1) were consistently upregulated greater than 3-fold. The 12 consistently upregulated genes also included CD59, polymerase (RNA) I polypeptide C, pyrophosphatase (inorganic) 1, mannose-P-dolichol utilization defect 1, nascent polypeptide-associated complex subunit alpha, hemoglobin-beta, fetuin-B, glyoxalase domain containing 4, and putative histone H3. The 28 upregulated ESTs represent genes with putative functions in the following five major categories: (1) immune response (46%); (2) signal transduction (21%); (3) transcriptional regulation (11%); (4) cell maintenance (11%); and (5) unknown (11%).