Download Differential gene expression profiling in healthy and white spot

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. Dhar1,2, Kate S. Licon1, Refugio Robles-Sikisaka1, Xiayou Zhang3, Robert A.
Bullis2 and Betsy Read4
1
Department of Biology, San Diego State University, 5500 Campanile Drive, San Diego,
California 92182, USA.
2
Advanced BioNutrition Corporation, 6430 Dobbin Road, Columbia, MD 21045.
3
Department of Computer Sciences, College of Arts and Sciences, California State
University, San Marcos, CA 92096.
4
Department of Biology, College of Arts and Sciences, California State University, San
Marcos, CA 92096.
*Corresponding author: Arun K. Dhar, Department of Biology, San Diego State
University, 5500 Campanile Drive, San Diego, California 92182, USA; Phone: 619-5944356; Fax: 619-594-5676; E-mail: [email protected] or [email protected]
Abstract
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.