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日本沼虾两种不同类型凝集素的分子克隆、特性分析及表达分析

Molecular cloning, characterization, and expression analysis of two different types of lectins from the oriental river prawn, Macrobrachium nipponense.

作者信息

Xiu Yunji, Wang Yinghui, Jing Yunting, Qi Yakun, Ding Zhengfeng, Meng Qingguo, Wang Wen

机构信息

Jiangsu Key Laboratory for Biodiversity & Biotechnology, College of Life Sciences, Nanjing Normal University, 1 Wenyuan Road, Nanjing 210046, China; Jiangsu Key Laboratory for Aquatic Crustacean Diseases, College of Life Sciences, Nanjing Normal University, 1 Wenyuan Road, Nanjing 210046, China.

Jiangsu Key Laboratory for Biodiversity & Biotechnology, College of Life Sciences, Nanjing Normal University, 1 Wenyuan Road, Nanjing 210046, China.

出版信息

Fish Shellfish Immunol. 2015 Aug;45(2):465-9. doi: 10.1016/j.fsi.2015.04.022. Epub 2015 Apr 28.

Abstract

Lectins, which are widely expressed in invertebrates, play important roles in many biological processes, including protein trafficking, cell signaling, pathogen recognition, as effector molecules, and so on (Wang and Wang, 2013). This study identified one novel M-type lectin and one L-Type lectin, designated as MnMTL1 and MnLTL1, from the oriental river prawn Macrobrachium nipponense. The full-length cDNA of MnMTL1 was 2064 bp with a 1761 bp ORF encoding a putative protein of 586 deduced amino acids. The full-length cDNA of MnLTL1 was 1744 bp with a 972 bp ORF encoding a 323-amino acid peptide. The deduced MnMTL1 protein contained a putative type II transmembrane region and a 440-aa Glycoside hydrolase family 47 (GH47) domain. One luminal carbohydrate recognition domain and a 23-aa type I transmembrane region were identified from the MnLTL1. MnMTL1 shared 78% identity with Marsupenaeus japonicus M-type lectin and MnLTL1 shared 83% similarity with M. japonicus L-type lectin. RT-PCR analysis showed that MnMTL1 and MnLTL1 were expressed in all tested tissues. Quantitative real-time PCR analysis revealed that MnMTL1 and MnLTL1 are substantially fluctuant during Aeromonas hydrophila and Aeromonas veronii infections. Based on immune responses and previous literature, we assumed that MnMTL1 and MnLTL1 might be functioned as pattern recognition receptors and play important roles in the immune response of M. nipponense.

摘要

凝集素在无脊椎动物中广泛表达,在许多生物过程中发挥重要作用,包括蛋白质运输、细胞信号传导、病原体识别、作为效应分子等(Wang和Wang,2013年)。本研究从日本沼虾(Macrobrachium nipponense)中鉴定出一种新型M型凝集素和一种L型凝集素,分别命名为MnMTL1和MnLTL1。MnMTL1的全长cDNA为2064 bp,开放阅读框(ORF)为1761 bp,编码一个推定的含586个氨基酸的蛋白质。MnLTL1的全长cDNA为1744 bp,ORF为972 bp,编码一个含323个氨基酸的肽段。推定的MnMTL1蛋白包含一个推定的II型跨膜区域和一个440个氨基酸的糖苷水解酶家族47(GH47)结构域。从MnLTL1中鉴定出一个腔内碳水化合物识别结构域和一个23个氨基酸的I型跨膜区域。MnMTL1与日本对虾(Marsupenaeus japonicus)的M型凝集素具有78%的同一性,MnLTL1与日本对虾的L型凝集素具有83%的相似性。RT-PCR分析表明,MnMTL1和MnLTL1在所有测试组织中均有表达。实时定量PCR分析显示,在嗜水气单胞菌(Aeromonas hydrophila)和维氏气单胞菌(Aeromonas veronii)感染期间,MnMTL1和MnLTL1有显著波动。基于免疫反应和先前的文献,我们推测MnMTL1和MnLTL1可能作为模式识别受体,在日本沼虾的免疫反应中发挥重要作用。

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