Identification of differentially expressed genes in shrimp (Penaeus stylirostris) infected with White spot syndrome virus by cDNA microarrays
Identification of differentially expressed genes in shrimp (Penaeus stylirostris) infected with White spot syndrome virus by cDNA microarrays
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DOI：
10.1007/s00705-003-0172-z
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发表时间：
2003-12-01
影响因子：
2.7
通讯作者：
Read, B
中科院分区：
文献类型：
--
作者：
Dhar, AK;Dettori, A;Read, B
白斑综合征病毒（White spot syndrome virus， WSSV）是目前世界上感染对虾最重要的病毒病原体。尽管在描述WSSV基因组特征和开发检测方法方面取得了相当大的进展，但有关参与WSSV发病机制的宿主基因的信息有限。我们研究了cDNA微阵列分析在研究wssv感染虾的基因表达方面的潜力。将虾的cdna以低密度阵列形式打印在玻片上，并与从健康虾和感染wssv的虾分离的RNA中获得的Cy3/Cy5标记探针杂交。感染WSSV后，与甲壳类动物免疫相关的蛋白、结构蛋白以及功能未知的蛋白的mRNA表达发生了改变。为了验证微阵列数据，通过实时RT-PCR检测了健康和感染wssv的对虾中三个差异表达基因的时间表达，一个免疫基因（c型凝集素-1），一个结构基因（40S核糖体蛋白）和一个脂质代谢基因（脂肪酸结合蛋白）。这些数据表明，WSSV感染改变了多种细胞基因的表达，并为进一步研究旨在鉴定其功能的基因提供了框架，这些基因可能有助于深入了解虾WSSV感染的机制。
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.