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源自抑制性消减杂交技术的鳜鱼感染传染性脾肾坏死病毒后脾脏中的差异基因表达谱

Differential gene expression profile in spleen of mandarin fish Siniperca chuatsi infected with ISKNV, derived from suppression subtractive hybridization.

作者信息

He Wei, Yinlt Zhi-Xin, Li You, Huo Wei-Li, Guan Hao-Ji, Weng Shao-Ping, Chan Siu-Ming, He Jian-Guo

机构信息

State Key Laboratory for Biocontrol, School of Life Sciences, Zhongshan University, 135 Xingang Road West, Guangzhou 510275, PR China.

出版信息

Dis Aquat Organ. 2006 Dec 14;73(2):113-22. doi: 10.3354/dao073113.

Abstract

To study the interaction between an invading virus and its host, we investigated differential gene expression in mandarin fish Siniperca chuatsi experimentally infected with infectious spleen and kidney necrosis virus (ISKNV). Subtractive cDNA libraries were constructed by suppression subtractive hybridization (SSH) from spleens of mock- and ISKNV-infected fish. Both forward- and reverse-subtracted libraries were generated. In the forward library, genes of the ubiquitin-proteasome proteolytic pathway, defense-related genes, a cytoskeletal protein gene, an apoptosis-related gene encoding inhibitor of apoptosis protein and JFC/EBPb cDNA for CAAT/Enhancer binding protein beta were up-regulated after infection. In the reverse library, genes that encoded CD59/neurotoxin/Ly-6-like protein, carboxypeptidase A2, and goose-type lysozyme were down-regulated. Some of these genes were analyzed by reverse transcription-polymerase chain reaction to confirm their differential expression as a result of virus infection. The results of this study may contribute to our understanding of fish innate immune response to ISKNV.

摘要

为了研究入侵病毒与其宿主之间的相互作用,我们调查了实验感染传染性脾肾坏死病毒(ISKNV)的鳜鱼(Siniperca chuatsi)的差异基因表达。通过抑制性消减杂交(SSH)从 mock 感染和 ISKNV 感染的鱼的脾脏构建了消减 cDNA 文库。生成了正向和反向消减文库。在正向文库中,泛素 - 蛋白酶体蛋白水解途径的基因、防御相关基因、一种细胞骨架蛋白基因、一种编码凋亡抑制蛋白的凋亡相关基因以及用于 CAAT/增强子结合蛋白β的 JFC/EBPb cDNA 在感染后上调。在反向文库中,编码 CD59/神经毒素/Ly - 6 样蛋白、羧肽酶 A2 和鹅型溶菌酶的基因下调。通过逆转录 - 聚合酶链反应对其中一些基因进行了分析,以确认它们因病毒感染而产生的差异表达。本研究结果可能有助于我们了解鱼类对 ISKNV 的先天免疫反应。

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