Characterization and expression analysis of an intelectin gene from Megalobrama amblycephala with excellent bacterial binding and agglutination activity
Characterization and expression analysis of an intelectin gene from Megalobrama amblycephala with excellent bacterial binding and agglutination activity
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团头鲂具有优异细菌结合和凝集活性的智力素基因的表征和表达分析
DOI：
10.1016/j.fsi.2016.12.023
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发表时间：
2017-02-01
影响因子：
4.7
通讯作者：
Liu, Hong
中科院分区：
文献类型：
--
作者：
Ding, Zhujin;Zhao, Xiaoheng;Liu, Hong
肠凝集素是近年来发现的一种凝集素，在机体的先天免疫、铁代谢和早期胚胎发育中起重要作用。intelectin在哺乳动物和两栖动物中的结构、表达模式和功能已被广泛研究，而在鱼类中的研究尚不清楚。本研究从团头鲂（Megalobrama amblycephala）中克隆了一个intelectin（MamINTL）基因，分析了其表达模式，并探讨了其在天然免疫应答中的作用。MamINTL cDNA编码312个氨基酸，前蛋白分子量为34 kDa。序列分析表明，在MamINTL中存在纤维蛋白原相关结构域和8个保守的半胱氨酸残基。MamINTL mRNA在不同发育阶段均可检测到，但在孵化后显著增加。在健康成年M.在团头菌中，MamINTL在各种组织中检测到，其中肝脏表达最高。在用嗜水气单胞菌攻击后，在肝、脾、肾、肠和鳃中观察到MamINTL mRNA的显著上调表达。此外，通过Western Blotting检测到的MamINTL蛋白水平的增加也在肝、肾和脾中观察到，表明MamINTL参与免疫应答。免疫组化分析M.团头虫肝脏切片显示感染后表达和定位的显著变化。此外，重组MamINTL对GFP表达的E.大肠杆菌中的Ca2+依赖性。本研究为深入了解鱼类智能素的特性、表达模式和功能提供了线索。(C)2016爱思唯尔有限公司版权所有。
Intelectin is a recently discovered lectin that plays vital roles in the innate immune response, iron metabolism and early embryogenesis. The structure, expression pattern and function of intelectin in mammals and amphibians have been well studied, while not well known in fish. In this study, we cloned a intelectin (MamINTL) gene from blunt snout bream (Megalobrama amblycephala), examined its expression patterns and explored its roles in innate immune response. The MamINTL cDNA encoded 312 amino acids, with a pro-protein of 34 kDa. Sequence analysis revealed the presence of a fibrinogen related domain and eight conserved cysteine residues in the MamINTL. The MamINTL mRNA was detectable at various developmental stages, while it increased significantly post hatching. In healthy adult M. amblycephala, MamINTL was detected in various tissues with the highest expression in the liver. Upon challenge with Aeromonas hydrophila, significantly up-regulated expression of the MamINTL mRNA was observed in the liver, spleen, kidney, intestine and gill. In addition, increased level of MamINTL protein detected by Western Blotting was also observed in the liver, kidney and spleen, indicating the participation of MamINTL in the immune response. Immunohistochemistry analysis of the M. amblycephala liver sections showed significant changes in expression and location post infection. In addition, the recombinant MamINTL showed excellent binding and agglutination activity against GFP-expressed E. coli in a Ca2+-dependent manner. Generally, the present study provides clues for a better understanding of the characterization, expression patterns and functions of fish intelectins. (C) 2016 Elsevier Ltd. All rights reserved.