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Gene expression profiling in gill tissues of White spot syndrome virus infected black tiger shrimp Penaeus monodon by DNA microarray

机译:基因芯片在白斑综合症病毒感染的黑虎虾斑节对虾g组织中的基因表达谱分析

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摘要

White spot syndrome virus (WSSV) continues to be the most devastating viral pathogen infecting penaeid shrimp the world over. The genome of WSSV has been deciphered and characterized from three geographical isolates and significant progress has been made in developing various molecular diagnostic methods to detect the virus. However, the information on host immune gene response to WSSV pathogenesis is limited. Microarray analysis was carried out as an approach to analyse the gene expression in black tiger shrimp Penaeus monodon in response to WSSV infection. Gill tissues collected from the WSSV infected shrimp at 6, 24, 48 h and moribund stage were analysed for differential gene expression. Shrimp cDNAs of 40,059 unique sequences were considered for designing the microarray chip. The Cy3-labeled cRNA derived from healthy and WSSV-infected shrimp was subjected to hybridization with all the DNA spots in the microarray which revealed 8,633 and 11,147 as up- and down-regulated genes respectively at different time intervals post infection. The altered expression of these numerous genes represented diverse functions such as immune response, osmoregulation, apoptosis, nucleic acid binding, energy and metabolism, signal transduction, stress response and molting. The changes in gene expression profiles observed by microarray analysis provides molecular insights and framework of genes which are up- and down-regulated at different time intervals during WSSV infection in shrimp. The microarray data was validated by Real Time analysis of four differentially expressed genes involved in apoptosis (translationally controlled tumor protein, inhibitor of apoptosis protein, ubiquitin conjugated enzyme E2 and caspase) for gene expression levels. The role of apoptosis related genes in WSSV infected shrimp is discussed herein.Electronic supplementary materialThe online version of this article (doi:10.1007/s13337-014-0243-7) contains supplementary material, which is available to authorized users.
机译:白斑综合症病毒(WSSV)仍然是全世界对虾的最具破坏力的病毒病原体。 WSSV的基因组已从三个地理分离株中破译并鉴定了特征,并且在开发检测病毒的各种分子诊断方法方面已经取得了重大进展。但是,有关宿主免疫基因对WSSV发病机理的信息有限。进行微阵列分析是分析黑虎虾对虾WSWS感染中基因表达的一种方法。在6、24、48小时和垂死阶段从WSSV感染的虾中收集的组织进行了差异基因表达分析。设计微阵列芯片时考虑了40,059个独特序列的虾cDNA。将来源于健康和被WSSV感染的虾的Cy3标记的cRNA与微阵列中的所有DNA斑点进行杂交,在感染后的不同时间间隔分别揭示了8,633和11,147个上调和下调的基因。这些众多基因表达的改变代表了多种功能,例如免疫应答,渗透调节,细胞凋亡,核酸结合,能量和代谢,信号转导,应激反应和蜕皮。通过微阵列分析观察到的基因表达谱的变化提供了对虾的WSSV感染期间不同时间间隔上调和下调的分子见解和基因框架。通过实时分析涉及细胞凋亡的四个差异表达基因(翻译控制的肿瘤蛋白,细胞凋亡蛋白的抑制剂,泛素结合酶E2和胱天蛋白酶),验证了微阵列数据的基因表达水平。凋亡相关基因在WSSV感染的虾中的作用将在本文中讨论。电子补充材料本文的在线版本(doi:10.1007 / s13337-014-0243-7)包含补充材料,授权用户可以使用。

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