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结合抑制性消减杂交和差异杂交技术分析抗白斑综合征病毒日本对虾血细胞中基因的差异表达谱

Differential profile of genes expressed in hemocytes of White Spot Syndrome Virus-resistant shrimp (Penaeus japonicus) by combining suppression subtractive hybridization and differential hybridization.

作者信息

He Nanhai, Qin Qiwei, Xu Xun

机构信息

Key Laboratory of Marine Biogenetic Resources, SOA, The Third Institute of Oceanography, State Oceanic Administration (SOA), 184# DaXue Road, Xiamen 361005, PR China.

出版信息

Antiviral Res. 2005 Apr;66(1):39-45. doi: 10.1016/j.antiviral.2004.12.010.

Abstract

White Spot Syndrome Virus (WSSV) is the major viral pathogen of culture shrimp. Although remarkable progress has been made in characterizing the WSSV genome, information concerning the antiviral process of host is still limited. To identify the genes differentially expressed along with their expression profile in the hemocytes of the virus-resistant shrimp, suppression subtractive hybridization (SSH) and differential hybridization (DH) were employed. Relying on the sequences identified in the subtractive cDNA library, 30 genes were characterized to be involved in the antiviral process as defense-relevant, among them, 22 are found for the first time in penaeid shrimp. The most interesting finding is that the interferon-like protein (IntlP) and (2'-5') oligo(A) synthetase-like protein (data not shown) known as the antiviral factors showed increased expression in virus-resistant shrimp and the non-specific antiviral activity of IntlP protein was verified by cytotoxicity experiment. A number of proteins with certain similarities to the components of the complement and cytokines system in vertebrates were also found in the subtracted library. The high expression of redox-related factors (NADH dehydrogenase, glutathione peroxidase and transcription factor AP-1 precursor), plasma defensive protein (C-type lectin and laminin-like protein) and translationally controlled tumor protein (TCTP) in the virus-resistant shrimp suggested that they are essential components participating in the antiviral process. Our work provides a wide array of genes differentially expressed in the virus-resistant shrimp, and a framework for further studies aimed at antiviral mechanism in shrimp.

摘要

白斑综合征病毒(WSSV)是养殖对虾的主要病毒病原体。尽管在WSSV基因组特征研究方面已取得显著进展,但关于宿主抗病毒过程的信息仍然有限。为了鉴定抗病毒对虾血细胞中差异表达的基因及其表达谱,采用了抑制性消减杂交(SSH)和差异杂交(DH)技术。基于消减cDNA文库中鉴定出的序列,有30个基因被确定参与抗病毒过程,属于防御相关基因,其中22个是首次在对虾中发现。最有趣的发现是,作为抗病毒因子的干扰素样蛋白(IntlP)和(2'-5')寡聚腺苷酸合成酶样蛋白(数据未显示)在抗病毒对虾中表达增加,并且通过细胞毒性实验验证了IntlP蛋白的非特异性抗病毒活性。在消减文库中还发现了一些与脊椎动物补体和细胞因子系统成分有一定相似性的蛋白质。抗病毒对虾中氧化还原相关因子(NADH脱氢酶、谷胱甘肽过氧化物酶和转录因子AP-1前体)、血浆防御蛋白(C型凝集素和层粘连蛋白样蛋白)以及翻译调控肿瘤蛋白(TCTP)的高表达表明它们是参与抗病毒过程的重要成分。我们的工作提供了一系列在抗病毒对虾中差异表达的基因,以及一个针对对虾抗病毒机制的进一步研究框架。

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