| 苏丹鱼甘露糖受体的基因克隆表达和免疫特性 |
| |
| 引用本文： | 何昕,秦真东,张凯,梁日深,杨森,伍剑标,赵丽娟,林蠡.苏丹鱼甘露糖受体的基因克隆表达和免疫特性[J].水产学报,2020,44(3):378-390. |
| |
| 作者姓名： | 何昕 秦真东 张凯 梁日深 杨森 伍剑标 赵丽娟 林蠡 |
| |
| 作者单位： | 广东省水环境与水产品安全工程技术研究中心,广州市水产病害与水禽养殖重点实验室,仲恺农业工程学院动物科技学院,广东广州 510225;佛山市南海区标记鱼苗场,广东佛山 528200 |
| |
| 基金项目： | 中国-东盟海上合作基金；国家自然科学基金（31872606，31572657，U1701233）；广东省海洋与渔业局基金（GDME-2018C006，D21822202，A201512C003，2015-115）；广东省教育厅基金（KA170500G，TK222001G，KA18058B3，KA1819604） |
| |
| 摘 要： | 为了解苏丹鱼MR(Lh MR)的结构特点及其在抗感染免疫反应中的作用,本研究克隆并分析了LhMR基因,采用实时荧光定量PCR(qRT-PCR)技术检测了LhMR在正常及柱状黄杆菌感染苏丹鱼组织中的表达。结果显示,LhMR开放阅读框4296 bp,编码1431个氨基酸(aa)。LhMR的氨基酸序列和分子结构与其他物种高度相似,胞外1个富含蓖麻类β型三叶草的结构域(RICIN)、1个纤连蛋白Ⅱ型结构域(FNⅡ)、8个串连的C型凝集素样结构域(CLECTs)、1个跨膜区和1个胞内区。通过qRT-PCR检测显示,LhMR在脾脏、体肾、心脏、脑、皮肤、肌肉、鳃、肝脏、后肠、前肠和中肠11种组织中均有表达,其中脾脏中表达量最高;经柱状黄杆菌感染苏丹鱼后,脾脏、肠、体肾和鳃中LhMR基因的相对表达量显著升高。通过免疫组织化学(IHC)检测发现,在脾脏、肾脏和鳃中的LhMR蛋白表达水平提高。通过苏木精-伊红染色病理组织切片表明,在体肾、肠、肝脏和鳃中均有不同程度的病变。体肾的肾小管有大量的血细胞浸润,上皮细胞发生坏死;肠壁变薄,组织大量弥散坏死;肝脏组织中有多个空泡;鳃小片肿胀、混乱、坏死和脱落。研究表明,LhMR在苏丹鱼感染柱状黄杆菌免疫反应中发挥重要作用,为苏丹鱼柱形病的防控提供新的参考。
|
| 关 键 词： | 苏丹鱼 柱状黄杆菌 甘露糖受体 克隆 基因表达 |
| 收稿时间： | 2019/2/26 0:00:00 |
| 修稿时间： | 2019/4/22 0:00:00 |
Cloning, expression and immune features of Sultan fish (Leptobarbus hoevenii) mannose receptor |
| |
| HE Xin,QIN Zhendong,ZHANG Kai,LIANG Rishen,YANG Sen,WU Jianbiao,ZHAO Lijuan and LIN Li.Cloning, expression and immune features of Sultan fish (Leptobarbus hoevenii) mannose receptor[J].Journal of Fisheries of China,2020,44(3):378-390. |
| |
| Authors: | HE Xin QIN Zhendong ZHANG Kai LIANG Rishen YANG Sen WU Jianbiao ZHAO Lijuan and LIN Li |
| |
| Institution: | Guangdong Provincial Water Environment and Aquatic Products Security Engineering Technology Research Center, Guangzhou Key Laboratory of Aquatic Animal Diseases and Waterfowl Breeding, Zhongkai University of Agriculture and Engineering, Guangzhou 510225, China,Guangdong Provincial Water Environment and Aquatic Products Security Engineering Technology Research Center, Guangzhou Key Laboratory of Aquatic Animal Diseases and Waterfowl Breeding, Zhongkai University of Agriculture and Engineering, Guangzhou 510225, China,Guangdong Provincial Water Environment and Aquatic Products Security Engineering Technology Research Center, Guangzhou Key Laboratory of Aquatic Animal Diseases and Waterfowl Breeding, Zhongkai University of Agriculture and Engineering, Guangzhou 510225, China,Guangdong Provincial Water Environment and Aquatic Products Security Engineering Technology Research Center, Guangzhou Key Laboratory of Aquatic Animal Diseases and Waterfowl Breeding, Zhongkai University of Agriculture and Engineering, Guangzhou 510225, China,Guangdong Provincial Water Environment and Aquatic Products Security Engineering Technology Research Center, Guangzhou Key Laboratory of Aquatic Animal Diseases and Waterfowl Breeding, Zhongkai University of Agriculture and Engineering, Guangzhou 510225, China,Biaoji Fishery Company, Nanhai District, Foshan 528200, China,Guangdong Provincial Water Environment and Aquatic Products Security Engineering Technology Research Center, Guangzhou Key Laboratory of Aquatic Animal Diseases and Waterfowl Breeding, Zhongkai University of Agriculture and Engineering, Guangzhou 510225, China and Guangdong Provincial Water Environment and Aquatic Products Security Engineering Technology Research Center, Guangzhou Key Laboratory of Aquatic Animal Diseases and Waterfowl Breeding, Zhongkai University of Agriculture and Engineering, Guangzhou 510225, China |
| |
| Abstract: | In order to understand the structural characteristics of MR and its role in anti-infective immune responses in Leptobarbus hoevenii, we cloned and sequenced the MR of L. hoevenii. Subsequently, real-time polymerase chain reaction (qRT-PCR) assay was used to measure the expression of the MR in the tissues of L. hoevenii with or without F. columnare infection. The results showed that the MR of L. hoevenii (LhMR) open reading frame was 4 296 bp, encoding 1 431 amino acids (aa). The amino acid sequence and molecular structure of LhMR are highly similar to the MRs from other animals, i.e., containing an extracellular ricin-like β-type clover domain (RICIN), a fibronectin type II domain (FNII) and eight tandem C Lectin-like domains (CLECTs), a transmembrane region and a short intracellular region. The mRNA of LhMR was widely expressed in 11 tested tissues, including spleen, kidney, heart, brain, skin, muscle, gills, liver, post intestine, fore intestine and mid intestine, with the highest expression level in the spleen. The mRNA expression of LhMR was significantly increased in the spleen, intestine, kidney and gills of L. hoevenii infected by F. columnare measured by qRT-PCR. Furthermore, the protein expression of LhMR was also increased in the spleen, kidney and gill of L. hoevenii infected by F. columnare revealed by immunochemistry assay. By Hematoxylin eosin staining, there were various pathological changes observed in the body kidney, intestine, liver and gills. In the body kidney, there were blood cells infiltrated in the renal tubules with necrosis in epithelial cells. The wall of intestine became thinner with diffused necrosis. Numerous vacuoles were observed in the liver cells. Gill lamella showed swollen, disordered, necrosis and detached. The results revealed that LhMR plays an important role in the immune responses of L. hoevenii infected by F. columnare and this will provide some reference for prevention and therapy strategies against columnaris disease. |
| |
| Keywords: | Leptobarbus hoevenii Flavobacterium columnare mannose receptor cloning gene expression |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ |
| 点击此处可从《水产学报》浏览原始摘要信息 |
| 点击此处可从《水产学报》下载免费的PDF全文 |
|
