草鱼钙网蛋白的克隆与组织表达  

Cloning and expression of Calreticulin full length cDNA in tissues of grass carp Ctenopharyngodon idella

作　　者：李旭威[1,2] 王海英[1] 吴锐全[1] 谢骏[1] 郁二蒙[1] 吴垠[2] 余德光[1] 王广军[1] 龚望宝[1] 

机构地区：[1]中国水产科学研究院 珠江水产研究所 农业部热带亚热带水产资源利用与养殖重点实验室,广东广州510380 [2]大连海洋大学水产与生命学院,辽宁大连116023

出　　处：《大连海洋大学学报》2012年第2期143-149,共7页Journal of Dalian Ocean University

基　　金：现代农业产业技术体系建设专项(nycytx-49);广东省海洋渔业科技推广项目(A200901D04);广东省自然科学基金资助项目(10151038001000004)

摘　　要：钙网蛋白是一种高度保守的分子伴侣蛋白,作为内质网主要的钙结合蛋白,广泛存在于真核生物细胞中,在病毒、寄生虫感染以及温度、氧气胁迫等情况下对细胞起保护作用。本试验中采用RACE方法首次克隆获得草鱼Ctenopharyngodon idellus钙网蛋白cDNA全序列,其全长为1 389 bp,开放阅读框为1 263bp,编码为421个氨基酸。将获得的草鱼钙网蛋白编码氨基酸序列与其它物种分别进行同源性比较,发现草鱼与斑马鱼Danio rerio、虹鳟Oncorhynchus mykiss的同源性较高,分别为86.26%、76.78%,而与人Homo sapiens、小家鼠Mus musculus的同源性为69.67%、69.19%。通过半定量RT-PCR检测可知,钙网蛋白在草鱼的肌肉、肠道、皮肤、肝胰脏、肾脏、脾脏、鳃和鳍中均有表达,其中在肝胰脏、鳍条中的表达量最高,除与皮肤中的表达量无显著差异外(P>0.05),均显著高于其它5个器官组织(P<0.05)。Calreticulin（CRT） as a highly conserved molecular chaperone protein extensively exists in eukaryotic cells,and CRT on the surface of the cell can protect cells in temperature,and oxygen stresses,pathogens and parasitic infections.The full length cDNA of calreticulin was cloned from grass carp Ctenopharyngodon idellus by race and pcr methods to ecvaluate the expression in different tissues.The gene was founf to consist of 1 389 bp with an open reading frame of 1 263 bp,encoding 421 amino acids.The amino acid sequence analysis showed that the grass carp Calreticulin had higher homology with zebra fish（86.26%）,and rainbow trout（76.78%）,and had 69.67% amino acid identity to murine and 69.19% to human CRT＇s.Tissue expression studies showed that CRT gene was all detected but expression was different in the tested tissues including muscle,skin,intestine,hepatopancreas,kidney,spleen,gills and rays,but the expression in the hepatopancreas liver and fins was found maximum,significantly higher than in the other five tissues except in skin（P0.05）.

关 键 词：草鱼 钙网蛋白 克隆 表达 2011-05-05 

分 类 号：S965.112[农业科学—水产养殖]