Differential profiles of gene expression in grouper Epinephelus coioides, infected with Singapore grouper iridovirus, revealed by suppression subtractive hybridization and DNA microarray.
Differential profiles of gene expression in grouper Epinephelus coioides, infected with Singapore grouper iridovirus, revealed by suppression subtractive hybridization and DNA microarray.
复制标题
DOI：
10.1111/j.1095-8649.2010.02676.x
复制
发表时间：
2010-08
影响因子：
2
通讯作者：
Dan Xu;Jingguang Wei;Huachun Cui;Jun Gong;Yang Yan;R. Lai;Qiwei Qin
中科院分区：
文献类型：
--
作者：
Dan Xu;Jingguang Wei;Huachun Cui;Jun Gong;Yang Yan;R. Lai;Qiwei Qin
使用抑制消减杂交 (SSH) 生成消减 cDNA 文库，该文库富含在感染新加坡石斑鱼虹彩病毒 (SGIV) 5 天后在橙斑石斑鱼脾脏中差异表达的基因转录本。在正向和反向消减文库中，分别鉴定了 260 个和 153 个非冗余表达序列标签 (EST)。这些注释的响应 SGIV 感染的基因被分为与免疫、细胞结构、转录翻译、细胞信号传导、代谢、线粒体蛋白、核糖体蛋白和未知或假设蛋白相关的八个基因类别。构建了包含所有差异表达基因的DNA微阵列，并研究了病毒感染的斜纹大肠杆菌不同组织中的基因表达模式。在这些基因中，有四个与感染过程相关的基因被鉴定出来，并通过定量实时 PCR 进行了进一步研究。这些结果为斜带大肠杆菌宿主-病原体相互作用的分子基础提供了新的见解，并将有助于制定针对 SGIV 感染的控制策略。
Suppression subtractive hybridization (SSH) was used to generate a subtracted cDNA library enriched with gene transcripts differentially expressed in the spleen of orange-spotted grouper Epinephelus coioides after 5 days of infection with Singapore grouper iridovirus (SGIV). In the forward and reverse-subtracted libraries, 260 and 153 non-redundant expressed sequence tags (EST), respectively, were identified. These annotated genes responding to SGIV infection were grouped into eight gene categories related to immunity, cell structure, transcription-translation, cell signalling, metabolism, mitochondrial proteins, ribosomal proteins and unknown or hypothetical proteins. A DNA microarray containing all the differentially expressed genes was constructed, and the gene expression patterns in different tissues were investigated in virus-infected E. coioides. Of these genes, four associated with the infection processes were identified and further investigated by quantitative real-time PCR. These results provide new insights into the molecular basis of host-pathogen interactions in E. coioides, and will help the development of control strategies against SGIV infection.