太平洋鳕IRF3 基因的克隆及绝对定量表达分析
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大连海洋大学 农业部北方海水增养殖重点实验室, 辽宁 大连 116023

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作者简介: 孙航(1990–), 男, 硕士研究生, 主要从事水产动物繁育研究. E-mail: sunhang1016@163.com 通信作者: 毛明光(1982–), 男, 博士, 讲师, 主要从事鱼类分子免疫学的研究. E-mail: mmg@dlou.edu.cn

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S917

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国家自然科学基金项目(31302202); 国家863 计划项目(2012AA10A413); 辽宁省教育厅项目(L2013276).


Cloning and expression analysis of IRF3 in Gadus macrocephalus usingabsolute quantitative PCR
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Key Laboratory of Mariculture Stock Enhancement in North China’s Sea, Ministry of Agriculture, Dalian Ocean University, Dalian 116023, China

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    摘要:

    3 (interferon regulatory factor 3,IRF3)(open reading frame, ORF), , 端(IRF association domain,IAD); (days post-hatching, dph)并对干扰素在仔鱼期的免疫作用进行分析。组织表达绝对定量结果显示肝和胸腺表达量高于其他组织。不同日龄仔鱼, 表达量大幅提高。本研究的结果为进一步研究太平洋鳕干扰素作用机制以及其在早期发育中的作用奠定了基础。

    Abstract:

    is a marine teleost that is economically important throughout the world. The specieshas a high nutritional value and is processed into a variety of products. As a result of the increase in fisheries targetingthis species, the abundance of wild populations has declined. Our laboratory has successfully carried out artificialbreeding and rearing of However, we observed significant mortality as a result of immune systemdysfunction. To improve artificial breeding techniques for , the molecular mechanisms and expressioncharacteristics of immune-related genes needs to be understood. Interferon regulatory factor 3 (IRF3) is a member ofthe interferon regulatory factor family. It is an important transcription factor for the expression of the interferon α/βgene and plays an important role in the host antiviral response mechanism. We obtained a cDNA sequence of IRF3 gene for the first time using gene cloning. The sequence was 1878 bp in length with a whole openreading frame (ORF) of 1377 bp that encoded a 459 amino acid protein. The molecular weight of the encoded proteinwas ~51 kDa. Amino acid sequence alignment revealed that the protein included a DNA binding domain (DBD) containingfive conserved tryptophan residues and a conserved C-terminal IRF association domain (IAD). The phylogenetictree revealed that the IRF3 of was clustered with the IRF3 of other species, distant from IRF1 andIRF2. The tissue and age-specific expression of IRF3 was detected using absolute quantitative PCR. Additionallytheeffect of interferon during the larval stages was analyzed. The expression levels were highest in the gonad, liver, andthymus. The copy number of IRF3 in the different tissues was: 231.244 copies/ng, 516.649 copies/ng, 695.158 copies/ng, 2128.273 copies/ng, 198.548 copies/ng, 101.758 copies/ng, 419.927 copies/ng, 13.016 copies/ng, 1102.775 copies/ng, and 13.016 copies/ng in the intestine, heart, thymus, gonad, gill, muscle, spleen, brain, liver, and kidney, respectively.The expression of IRF3 was detected in fertilized eggs and remained relatively constant to 5 d post-hatching (dph)but increased by 25 dph. The copy number of IRF3 at different ages was: 6.189 copies/ng, 6.809 copies/ng, 8.066 copies/ng, 5.009 copies/ng, 5.009 copies/ng, 10.390 copies/ng, and 10.390 copies/ng in the fertilized egg, 1 dph, 5 dph, 9dph, 17 dph, 25 dph, or 33 dph, respectively. Our results suggest that the gonad, liver, and thymus are the primary organsfor IRF3 expression. Additionally, our results suggest that interferon plays an antiviral role at 25 dph. The resultsof this study lay the foundation for further study of the mechanism of interferon action during the early development of

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孙航,姜志强,蒋洁兰,温施慧,李幸,暴宁,苏鹏,毛明光.太平洋鳕IRF3 基因的克隆及绝对定量表达分析[J].中国水产科学,2015,22(5):887-895
SUN Hang, JIANG Zhiqiang, JIANG Jielan, WEN Shihui, LI Xing, BAO Ning, SU Peng, MAO Mingguang. Cloning and expression analysis of IRF3 in Gadus macrocephalus usingabsolute quantitative PCR[J]. Journal of Fishery Sciences of China,2015,22(5):887-895

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  • 在线发布日期: 2015-09-15
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