团头鲂黏蛋白基因Muc5b克隆及表达分析  

Cloning and Expression of Muc5b in Megalobrama amblycephala and Muc5b Expression Alteration after Catching Stress

作　　者：薛春雨[1,2] 董晶晶[1] 习丙文[2,3] 任鸣春[2] 梁利国[2] 谢骏[1,2] 

机构地区：[1]南京农业大学无锡渔业学院,无锡214081 [2]中国水产科学研究院淡水渔业研究中心农业部淡水渔业和种质资源利用重点实验室,无锡214081 [3]淡水水产健康养殖湖北省协同创新中心,武汉430072

出　　处：《动物学杂志》2014年第6期886-896,共11页Chinese Journal of Zoology

基　　金：现代农业产业技术体系建设专项(No.CARS-46-10);中央级公益性科研院所基本科研业务费专项资金项目(No.2013JBFM10)

摘　　要：黏液(mucus)在鱼体防御外界病原侵袭、信息传递、调节渗透压等方面具有重要作用。黏蛋白(mucin)作为黏液的基础骨架组分,与其相关的研究正受到广泛的关注。在本研究中,作者克隆获得团头鲂(Megalobrama amblycephala)Muc5b mRNA的部分序列3 895 bp,并通过qRT-PCR分析了Muc5b在团头鲂不同组织的表达分布及其在捕捞应激后在鳃和表皮中的表达变化。序列分析结果显示,团头鲂Muc5b与鲤(Cyprinus carpio)等脊椎动物的Muc5b有较高的同源性,其N端含有黏蛋白特异性结构域:3个VWD区域,3个C8区域,2个TIL区。组织表达分析结果表明,Muc5b在鳃和表皮表达量相对较高,在脑、脾、肾中表达水平较低,在肝、肠道几乎不表达。捕捞应激后1 h时鳃中Muc5b显著降低(P<0.05),24 h时恢复初始水平;表皮中Muc5b在4 h时显著上升(P<0.05),24 h时恢复到初始水平。Mucus plays an important role in protecting fish against pathogen invasion,signaling events,regulating osmotic pressure and so on. As mucins are the important skeleton components of the mucus,the research on fish mucins has caught wide attention. However,few studies have been reported on fish mucin gene characteristics and expression pattern. In this study,Megalobrama amblycephala Muc5 b mRNA（ 3 895 bp length） was cloned and its tissue expression was analyzed by qRT-PCR. The result showed that sequence of M.amblycephala Muc5 b had high homology with that of other vertebrates,and its N-terminal of predicted amino acid sequence contained several non-PTS region domains： three VWD domains,three C8 domains and two TIL domains（ Fig. 2）. Gene expression analysis showed relatively higher expression of Muc5 b in gills and skin,lower expression in brain,spleen,kidney,and almost no expression in liver and intestine（ Fig. 4）. Inaddition,variation in Muc5 b expression was detected in gills and skin after catching stress. Muc5 b expression was significantly reduced in the gills（ t-test,P 〈 0. 05） at 1 h after catching-stress,and recovered to the initial level at 24 h; On the contrary,Muc5 b expression was increased in the skin at 4 h（ t-test,P 〈 0. 05）,then recovered to the initial level（ Fig. 6）. Thus,the Muc5 b expression can be influenced by catching-stress.

关 键 词：团头鲂 黏液 Muc5b 应激 

分 类 号：S917.4[农业科学—水产科学]