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United States Department of Agriculture

Agricultural Research Service

Research Project: MOLECULAR ANALYSIS OF VIRULENCE DETERMINANTS OF SELECT BACTERIA IN FISH DISEASES

Location: Aquatic Animal Health Research

Title: Identification and expression profile of multiple genes in the anterior kidney of channel catfish induced by modified live Edwardsiella ictaluri vaccination.

Authors
item Wei Pridgeon, Yuping
item Shoemaker, Craig
item Klesius, Phillip

Submitted to: Veterinary Immunology and Immunopathology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: September 13, 2009
Publication Date: March 1, 2010
Repository URL: http://hdl.handle.net/10113/41644
Citation: Pridgeon, Y.W., 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. Veterinary Immunology and Immunopathology. 134:184-198.

Interpretive Summary: To understand the molecular responses of channel catfish to modified live vaccine of bacterium Edwardsiella ictaluri, PCR-select subtractive cDNA hybridization technique was used to isolate genes that are induced by the vaccine. A total of 124 clones were isolated from a subtractive library of modified live Edwardsiella ictaluri-vaccinated vs sham-vaccinated channel catfish anterior kidney. A total of 57 different genes were obtained from the 240 clones. The transcription levels of the 57 genes in response to E. ictaluri vaccination were then evaluated by quantitative PCR (QPCR). Of the 57 ESTs, 43 were induced/upregulated at least 2 fold higher in all three vaccinated fish compared to unvaccinated control fish. Of the 43 upregulated genes, five were consistently upregulated greater than 10 fold, including two highly upregulated (>20 fold) glycosyltransferase and Toll-like receptor 5. Three genes (GTPase 1, coatomer protein complex zeta 1, and type II arginine deiminase) were consistently induced greater than 10 fold. Two genes (MHC class I alpha chain and transposase) were upregulated greater than 10 fold in two of the three vaccinated fish. The 43 upregulated genes also included 19 moderately upregulated (3 to 10 fold) and 17 slightly upregulated (2 to 3 fold). 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.

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). Of the 57 ESTs, 43 were induced at least 2 fold higher in all three vaccinated fish compared to unvaccinated control fish. Of the 43 upregulated genes, five were consistently upregulated greater than 10 fold, including two highly upregulated (>20 fold) glycosyltransferase and Toll-like receptor 5. The transcriptional levels of GTPase 1, coatomer protein complex zeta 1, and type II arginine deiminase were consistently induced greater than 10 fold. MHC class I alpha chain and transposase were upregulated greater than 10 fold in two of the three vaccinated fish. The 43 upregulated genes also included 19 moderately upregulated (3 to 10 fold) and 17 slightly upregulated (2 to 3 fold). 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.

Last Modified: 7/30/2014
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