Cloning and preliminary functional studies of the JAM-A gene in grass carp (Ctenopharyngodon idellus)
Cloning and preliminary functional studies of the JAM-A gene in grass carp (Ctenopharyngodon idellus)
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DOI：
10.1016/j.fsi.2013.03.352
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发表时间：
2013-06-01
影响因子：
4.7
通讯作者：
Wang, Yaping
中科院分区：
文献类型：
--
作者：
Du, Fukuan;Su, Jianguo;Wang, Yaping
草鱼(Ctenophyngodon Idellus)是中国等东南亚国家非常重要的水产养殖物种，但该物种疾病暴发频繁，造成巨大的经济损失。草鱼呼肠孤病毒(GCRV)是由草鱼呼肠孤病毒(GCRV)引起的草鱼出血性疾病。连接黏附分子A(JAM-A)是呼肠孤病毒的哺乳动物受体，已被广泛研究。然而，到目前为止，对草鱼JAM-A基因的研究还很少。在本研究中，我们克隆并阐明了草鱼JAM-A基因(GcJAM-A)的结构，并对其在草鱼出血中的功能进行了研究。GcJAM-A由10个外显子和9个内含子组成，全长1833bp，含有888bp的开放阅读框，编码295个氨基酸的蛋白质。GcJAM-A蛋白被预测包含一个典型的跨膜区。GcJAM-A的母体表达模式在胚胎发育早期即可观察到，而合子则在孵化后8h开始表达。GcJAM-A在鳃、肝、肠和肾中表达较强，在血液、脑、脾和头肾中表达较弱。此外，与6月龄相比，30月龄个体的鳃、肝、肠、脑、脾和肾中的表达水平较低。在感染GCRV的GcJAM-A基因敲除细胞系(CIK)中，干扰素和细胞凋亡途径相关基因的表达受到显著抑制。这些结果提示GcJAM-A可能是GCRV的受体。因此，我们成功地确定了GcJAM-A基因的特征，并为其作为GCRV受体的作用提供了证据。皇冠版权所有(C)2013由爱思唯尔有限公司出版。保留所有权利。
Grass carp (Ctenopharyngodon idellus) is a very important aquaculture species in China and other South-East Asian countries; however, disease outbreaks in this species are frequent, resulting in huge economic losses. Grass carp hemorrhage caused by grass carp reovirus (GCRV) is one of the most serious diseases. Junction adhesion molecule A (JAM-A) is the mammalian receptor for reovirus, and has been well studied. However, the JAM-A gene in grass carp has not been studied so far. In this study, we cloned and elucidated the structure of the JAM-A gene in grass carp (GcJAM-A) and then studied its functions during grass carp hemorrhage. GcJAM-A is composed of 10 exons and 9 introns, and its full-length cDNA is 1833 bp long, with an 888 bp open reading frame (ORE) that encodes a 295 amino acid protein. The GcJAM-A protein is predicted to contain a typical transmembrane domain. Maternal expression pattern of GcJAM-A is observed during early embryogenesis, while zygote expression occurs at 8 h after hatching. GcJAM-A is expressed strongly in the gill, liver, intestine and kidney, while it is expressed poorly in the blood, brain, spleen and head kidney. Moreover, lower expression is observed in the gill, liver, intestine, brain, spleen and kidney of 30-month-old individuals, compared with 6-month-old. In a GcJAM-A-knockdown cell line (CIK) infected with GCRV, the expression of genes involved in the interferon and apoptosis pathways was significantly inhibited. These results suggest that GcJAM-A could be a receptor for GCRV. We have therefore managed to characterize the GcJAM-A gene and provide evidence for its role as a receptor for GCRV. Crown Copyright (C) 2013 Published by Elsevier Ltd. All rights reserved.