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抑制差减杂交(SSH)和实时 PCR 显示在太平洋牡蛎(Crassostrea gigas)中挑战疱疹病毒 1 后差异基因表达。

Suppression substractive hybridisation (SSH) and real time PCR reveal differential gene expression in the Pacific cupped oyster, Crassostrea gigas, challenged with Ostreid herpesvirus 1.

机构信息

Ifremer, Laboratoire de Génétique et Pathologie, 17390 La Tremblade, France.

出版信息

Dev Comp Immunol. 2011 Jul;35(7):725-35. doi: 10.1016/j.dci.2011.02.004. Epub 2011 Mar 1.

Abstract

Virus-induced genes were identified using suppression subtractive hybridisation (SSH) from Pacific cupped oyster, Crassostrea gigas, haemocytes challenged by OsHV-1. A total of 304 clones from SSH forward library were sequenced. Among these sequences, some homologues corresponded to (i) immune related genes (macrophage express protein, IK cytokine, interferon-induced protein 44 or multicopper oxidase), (ii) apoptosis related genes (Bcl-2) and (iii) cell signalling and virus receptor genes (glypican). Molecular characterization and phylogenic analysis of 3 immune-related genes (macrophage expressed protein, multicopper oxidase and immunoglobulin domain cell adhesion molecule) were performed. Finally, quantitative PCR revealed significant changes in the expression of immune related genes (multicopper oxidase, macrophage expressed protein, myeloid differentiation factor 88 and interferon-induced protein 44) in oysters experimentally challenged with OsHV-1. These findings provide a first basis for studying the role of innate immunity in response to viruses in bivalves and identified genes may serve as markers of interest in breeding programs in order to obtain selected oysters presenting OsHV-1 resistance.

摘要

利用抑制性消减杂交(SSH)技术,从太平洋牡蛎(Crassostrea gigas)血细胞中鉴定出病毒诱导基因,这些血细胞受到了 OsHV-1 的挑战。从 SSH 正向文库中总共测序了 304 个克隆。在这些序列中,一些同源物对应于(i)免疫相关基因(巨噬细胞表达蛋白、IK 细胞因子、干扰素诱导蛋白 44 或多铜氧化酶)、(ii)凋亡相关基因(Bcl-2)和(iii)细胞信号和病毒受体基因(糖蛋白)。对 3 种免疫相关基因(巨噬细胞表达蛋白、多铜氧化酶和免疫球蛋白结构域细胞黏附分子)进行了分子特征和系统发育分析。最后,定量 PCR 显示,在 OsHV-1 实验性感染的牡蛎中,免疫相关基因(多铜氧化酶、巨噬细胞表达蛋白、髓样分化因子 88 和干扰素诱导蛋白 44)的表达发生了显著变化。这些发现为研究贝类先天免疫对病毒反应的作用提供了第一个基础,并确定了一些可能作为选择具有 OsHV-1 抗性的牡蛎的育种计划中感兴趣的标记基因。

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