|Wei Pridgeon, Yuping|
Submitted to: World Aquaculture Society Meeting
Publication Type: Proceedings
Publication Acceptance Date: 11/30/2009
Publication Date: 3/1/2010
Citation: Wei Pridgeon, Y., Shoemaker, C.A., Klesius, P.H. 2010. Identification and expression profile of multiple genes in the anterior kidney of channel catfish induced by modified live Edwardsiella ictaluri vaccination. Proceedings of Aquaculture 2010. p. 795.
Technical Abstract: Using PCR-select subtractive cDNA hybridization technique, 57 expressed sequence tags (ESTs) were isolated from 240 clones of a modified live Edwardsiella ictaluri-vaccinated vs sham-vaccinated channel catfish anterior kidney subtractive library. The transcription levels of the 57 ESTs in response to E. ictaluri vaccination were then evaluated by quantitative PCR (qPCR). The 57 genes isolated from the subtractive library were classified in terms of their putative functions. Half of the genes identified were either involved in immune-response or metabolism (Figure 1). The major portion (28%) of the 57 genes identified were immune-related genes, including MHC class I alpha chain, surface antigen BspA-like protein/NK-lysin type 3, complement C4a, leukocyte cell-derived chemotaxin 2 (LECT2), CD83 antigen precursor, lysozyme g, CD45 precursor, and Toll-like receptor 5, glucose phosphate isomerase a, and TNF alpha induced protein 2 (TNFAIP2). Another major group (21%) of the identified genes was related to metabolism, NADH dehydrogenase subunit 3 and cytochrome b. Six genes (10%) were related to apoptosis, including huntingtin interacting protein K and ring finger 144B. Eight genes (14%) were related to cell growth and maintainance, including beta-actin and alpha-tubulin. Three genes (5%) were related to endocytosis and lysosome trafficking, including lysosomal-associated transmembrane protein 5 (LAMP5) and Ras-related protein Rab-11A. Three genes (5%) were related to gene regulation, including pre-Mrna processing factor 8 and eukaryotic translation initiation factor 3. One gene (2%), caldendrin, was involved in calcium signaling. The functions of five genes (9%) are currently unknown. Our results suggest that subtractive cDNA hybridization and qPCR are powerful cost-effective techniques to identify differentially expressed genes in response to modified live E. ictaluri vaccination.