Plant & Animal Genomes XIV Conference

January 14-18, 2006
Town & Country Convention Center
San Diego, CA

Differential Gene Expression Profiling In Healthy And White Spot Syndrome (WSSV) Virus- Infected Shrimp (Penaeus stylirostris) By EST Analysis Isolated By Suppression Subtractive Hybridization.

Arun K. Dhar^1,2 , Kate S. Licon¹ , Refugio Robles-Sikisaka¹ , Xiayou Zhang³ , Robert A. Bullis² , Betsy A. Read⁴

¹  Department of Biology, San Diego State University, 5500 Campanile Dr., San Diego, CA 92182, USA

²  Advanced BioNutrition Corporation, 6430 Dobbin Road, Columbia, MD 21045, USA

³  Department of Computer Sciences, College of Arts and Sciences, California State University, San Marcos, CA 92096, USA

⁴  Department of Biological Sciences, College of Arts and Sciences, California State University, San Marcos, CA 92096, USA

White spot syndrome disease, caused by the white spot syndrome virus (WSSV) is the most important viral disease of shrimp aquaculture worldwide. To identify genes involved in antiviral response, expressed sequence tags (ESTs) were isolated from hepatopancreas cDNA libraries of healthy and WSSV-infected shrimp made by reciprocal suppression subtractive hybridization. The efficacy of each of the libraries was validated using dot blot and virtual northern blot hybridizations. A total of 956 clones (478 clones from each of healthy and infected library) were sequenced. Seventy seven unigenes were obtained from the healthy library including 31 novel genes that showed no similarity with the GenBank database entries. From the WSSV-infected library, a total of 106 unigenes were obtained, 47 of which showed no similarity with database entries. Hemocyanin was the most abundant gene in both libraries. 19 different immune and/or defense related transcripts were detected in the library constructed from WSSV-infected shrimp whereas 8 immune and/or defense related genes were detected in the library constructed from healthy shrimp. Some of the more interesting immune related transcripts up-regulated in the virally infected shrimp include a high density lipoprotein/1,3-beta-D glucan binding protein, chitinase, lectin, zinc proteinase. Furthermore, a number of proteins potentially involved in signal transduction were identified in the WSSV-infected library. The temporal expression of a subset of the immune or defense related genes are currently underway. Functional analysis of these genes will contribute to our efforts to understand the molecular underpinnings of WSSV pathogenesis in shrimp.