斜带石斑鱼IFN-γ基因的克隆与表达分析
Molecular characterization and expression of interferon gamma(IFN-γ) in orange-spotted grouper(Epinephelus coioides)
查看参考文献22篇
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文摘
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IFN-γ在天然免疫和适应性免疫应答中均发挥着重要的作用。由于低等脊椎动物IFN-γ与哺乳动物IFN-γ相似性非常低,硬骨鱼类的IFN-γ近年来才得到鉴定。本研究报道了斜带石斑鱼(Epinephelus coioides)的IFN-γ基因,并对其表达进行了分析。斜带石斑鱼IFN-γcDNA全长为867 bp,编码200个氨基酸。其基因组全长为5 104 bp,由4个外显子、3个内含子组成。氨基酸序列比对显示斜带石斑鱼IFN-γ与高等脊椎动物IFN-γ的序列相似性较低,仅为32.5%~39%,与其他硬骨鱼类的IFN-γ相似性为38.0%~68.5%。序列分析结果显示,斜带石斑鱼IFN-γ分子C末端中存在IFN-γ的特征序列以及核定位基序。二级结构分析结果显示斜带石斑鱼IFN-γ序列中包含6个α螺旋结构,其排列与高等脊椎动物的IFN-γ类似。应用荧光定量PCR检测了IFN-γ基因在Poly I:C诱导后的表达变化。发现在肾、脾、鳃、头肾、皮肤和胸腺组织中,Poly I:C能显著诱导IFN-γ的表达,在胸腺中上调倍数最高,其次为头肾、脾、肾、皮肤和鳃,而在脑和心脏组织中没有显著变化。据此推测,IFN-γ在斜带石斑鱼抵抗病毒感染的过程中起有十分重要的作用。 |
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其他语种文摘
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IFN-γ is a typeⅡinterferon that plays an important role in both innate and cell-mediated immune responses. Because of the low similarity with their mammalian homologues, IFN-γ in teleost fish has only recently been identified. In this study, the cDNA sequence encoding the IFN-γ molecule was cloned from the orange-spotted grouper, Epinephelus coioides, and its gene organization was revealed. Its expression in response to Poly I:C stimulation was also examined. The IFN-γ cDNA sequence consists of 867 bp, which encodes a putative protein of 200 amino acids. The gene is 5-kb long and contains four exons, which are comparable in size with IFN-γ genes in other vertebrates. This IFN-γ was only 32.5%-39% similar to its counterparts in higher vertebrates and 38.0%-68.5% similar to those in other teleost fish. Sequence analysis revealed that the IFN-γ in the orange-spotted grouper contains the typical IFN-γ motif and a conserved nuclear localization site in the C-terminal end and displays an alpha-helix structure similar to mammalian IFN-γ. Real time PCR analysis results showed that the expression of IFN-γ increased in the thymus, head kidney, spleen, kidney, skin, and gills following the poly I:C stimulation, indicating its role against viruses in orange-spotted grouper. |
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来源
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中国水产科学
,2013,20(2):269-275 【核心库】
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DOI
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10.3724/sp.j.1118.2013.00269
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关键词
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Ⅱ型干扰素
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IFN-γ
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基因表达
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斜带石斑鱼
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地址
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1.
中国科学院水生生物研究所, 淡水生态与生物技术国家重点实验室, 湖北, 武汉, 430072
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集美大学水产学院, 福建, 厦门, 361021
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语种
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中文 |
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文献类型
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研究性论文 |
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ISSN
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1005-8737 |
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学科
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水产、渔业 |
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基金
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国家自然科学基金-广东联合基金
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国家863计划
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文献收藏号
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CSCD:4806345
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