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日本牙鲆干扰素调节因子 10(IRF10)的分子克隆与表达分析。

Molecular cloning and expression analysis of interferon regulatory factor 10 (IRF10) in Japanese flounder, Paralichthys olivaceus.

机构信息

Laboratory of Genome Science, Graduate School of Marine Science and Technology, Tokyo University of Marine Science and Technology, Konan 4-5-7, Minato, Tokyo 108-8477, Japan.

出版信息

Fish Shellfish Immunol. 2011 Jan;30(1):67-76. doi: 10.1016/j.fsi.2010.09.010. Epub 2010 Sep 29.

Abstract

Interferons (IFNs) are important for the defense against intracellular pathogens. Interferon regulatory factor 10 (IRF10) is a transcript factor involved in regulating IFN signaling induced by virus infections in birds, but little is know of its role in the immune response in non-avian vertebrates. Here, we identified and characterized the IRF10 gene and examined its expression pattern in a teleost fish, Japanese flounder, Paralichthys olivaceus. Japanese flounder IRF10 (PoIRF10) gene is a single copy gene, contains 8 exons and 7 introns and has 4918 nucleotides (nt) including an open reading frame (ORF) of 1212nt encoding 404 amino acids. The deduced PoIRF10 amino acid sequence contains a DNA-binding domain (DBD), nuclear localization signal (NLS) and IRF association domain (IAD). PoIRF10 is most closely related to chicken and zebrafish IRF10 by homology search and phylogenetic analysis. Putative binding sites for activator protein-1 (AP-1), CAAT-enhancer binding protein (C/EBP), C/EBPβ, cAMP response element-binding protein (CRE-BP), IFN-stimulated response element (ISRE) and signal transducer and activator of transcription (STAT) were found in the 5' flanking region (2.0kb) of PoIRF10 gene. PoIRF10 mRNA was strongly expressed in gill, head kidney, heart, peripheral blood lymphocytes (PBLs), spleen and trunk kidney. PoIRF10 expression was up-regulated by Edwardsiella tarda, Streptococcus iniae and viral hemorrhagic septicemia virus (VHSV) infection in kidney. Lipopolysaccharide (LPS) and poly I:C also increased PoIRF10 expression level in PBLs. These results suggest that PoIRF10 is related to immune response in not only virus infection but also bacterial infection in teleost fish and should help to clarify the biological function of IRF10.

摘要

干扰素(IFNs)对于抵抗细胞内病原体非常重要。干扰素调节因子 10(IRF10)是一种转录因子,参与调节鸟类病毒感染诱导的 IFN 信号通路,但在非鸟类脊椎动物中,其在免疫反应中的作用知之甚少。在这里,我们鉴定并表征了鱼类日本牙鲆的 IRF10 基因,并研究了其在日本牙鲆中的表达模式。日本牙鲆 IRF10(PoIRF10)基因是一个单拷贝基因,包含 8 个外显子和 7 个内含子,具有 4918 个核苷酸(nt),包括一个编码 404 个氨基酸的开放阅读框(ORF)。推导的 PoIRF10 氨基酸序列包含一个 DNA 结合域(DBD)、核定位信号(NLS)和 IRF 结合域(IAD)。通过同源搜索和系统发育分析,PoIRF10 与鸡和斑马鱼 IRF10 最为密切相关。在 PoIRF10 基因的 5'侧翼区(2.0kb)中发现了潜在的激活蛋白-1(AP-1)、CAAT 增强子结合蛋白(C/EBP)、C/EBPβ、cAMP 反应元件结合蛋白(CRE-BP)、干扰素刺激反应元件(ISRE)和信号转导和转录激活因子(STAT)结合位点。PoIRF10 mRNA 在鳃、头肾、心脏、外周血淋巴细胞(PBLs)、脾脏和躯干肾中强烈表达。在肾脏中,爱德华氏菌、链球菌和病毒性出血性败血症病毒(VHSV)感染可上调 PoIRF10 的表达。脂多糖(LPS)和聚 I:C 也可增加 PBLs 中 PoIRF10 的表达水平。这些结果表明,PoIRF10 不仅与病毒感染有关,而且与鱼类的细菌感染有关,这有助于阐明 IRF10 的生物学功能。

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