Dhar A K, Dettori A, Roux M M, Klimpel K R, Read B
Department of Biology, San Diego State University, 5500 Campanile Drive, San Diego, CA 92182, U.S.A.
Arch Virol. 2003 Dec;148(12):2381-96. doi: 10.1007/s00705-003-0172-z. Epub 2003 Oct 9.
White spot syndrome virus (WSSV) is currently the most important viral pathogen infecting penaeid shrimp worldwide. Although considerable progress has been made in characterizing the WSSV genome and developing detection methods, information pertaining to host genes involved in WSSV pathogenesis is limited. We examined the potential of cDNA microarray analysis to study gene expression in WSSV-infected shrimp. Shrimp cDNAs were printed as low-density arrays on glass slides and were hybridized with Cy3/Cy5 labeled probes derived from RNA isolated from healthy and WSSV-infected shrimp. Genes that code for proteins that are relevant to crustacean immunity, structural proteins, as well as proteins of unknown function were among those whose mRNA expression was altered upon WSSV infection. To validate the microarray data, the temporal expression of three differentially expressed genes, an immune gene (C-type lectin-1), a structural gene (40S ribosomal protein), and a gene involved in lipid metabolism (fatty acid binding protein) was measured in healthy and WSSV-infected shrimp by real-time RT-PCR. The data suggest that WSSV infection alters the expression of a wide array of cellular genes, and provides a framework for further studies aimed at identifying genes whose function may provide insight into the mechanism of WSSV infection in shrimp.
白斑综合征病毒（WSSV）是目前全球对虾感染中最重要的病毒病原体。尽管在WSSV基因组特征描述和检测方法开发方面取得了显著进展，但与WSSV发病机制中涉及的宿主基因相关的信息仍然有限。我们研究了cDNA微阵列分析在研究WSSV感染对虾基因表达方面的潜力。将对虾cDNA以低密度阵列形式点印在玻片上，并与从健康和WSSV感染对虾中分离的RNA衍生的Cy3/Cy5标记探针进行杂交。在WSSV感染后，其mRNA表达发生改变的基因包括编码与甲壳类免疫相关蛋白质、结构蛋白以及功能未知蛋白质的基因。为了验证微阵列数据，通过实时RT-PCR检测了健康和WSSV感染对虾中三个差异表达基因的时间表达情况，这三个基因分别是一个免疫基因（C型凝集素-1）、一个结构基因（40S核糖体蛋白）和一个参与脂质代谢的基因（脂肪酸结合蛋白）。数据表明，WSSV感染会改变多种细胞基因的表达，并为进一步研究提供了一个框架，旨在鉴定那些其功能可能有助于深入了解WSSV在对虾中感染机制的基因。
