草鱼瘦素受体基因片段序列的克隆及其组织表达分析  

Cloning and Tissue Expression Analysis of the Leptin Receptor Gene Fragment Sequence in Grass Carp(Ctenopharyngodon idella)

作　　者：吴小凤[1] 李小勤[1] 冷向军[1] 关磊[1] 郭婷[1] 

机构地区：[1]上海海洋大学省部共建水产种质资源发掘与利用教育部重点实验室,上海201306

出　　处：《生物技术通报》2011年第11期118-124,共7页Biotechnology Bulletin

基　　金：上海市教委发展基金(06KZ002);上海海洋大学"教育部水产种质资源发掘与利用省部共建重点实验室"及"农业部水产种质资源与利用重点开放实验室"开放课题基金(KFT2008-6);中国南海水产研究所种质资源开放课题(2008A003);上海市水产养殖重点学科建设项目(Y1101)

摘　　要：根据斑马鱼、大西洋鲑和人等物种已知的瘦素受体基因核苷酸保守区序列设计一对简并引物,通过RT-PCR法从草鱼肝胰脏中首次克隆获得草鱼瘦素受体基因的片段序列。该片段序列长713 bp,编码237个氨基酸,氨基酸序列分析表明草鱼瘦素受体基因片段氨基酸序列与其他物种的相似性在35%-86%之间。通过邻接法(Neighbor Joining,NJ)构建系统进化树显示,鱼类的瘦素受体独立聚成一支,草鱼与金鱼、斑马鱼聚成一支,再与日本青鳉、黑点青鳉、红鳍东方鲀和大西洋鲑聚成一支。通过实时荧光定量PCR分析草鱼瘦素受体基因的组织差异表达,结果表明,草鱼瘦素受体基因在肝胰脏、肌肉、脑、心脏、脾和肠系膜脂肪组织中均有表达,其中在脾脏组织中表达量最多,显著高于其他组织(P<0.05),其次是心脏、脑、肌肉和肠系膜脂肪组织,在肝胰脏组织中表达量最低,且显著低于其他组织(P<0.05)。According to zebrafish,Atlantic salmon,human and other species＇ leptin receptor gene conservative region of nucleotide sequences,degenerate oligonucleotide primer was designed and the leptin receptor gene fragment sequence was amplified by RT-PCR from liver of grass carp（Ctenopharyngodon idella）.The obtained cDNA fragment sequence was 713 bp,which encoded 237 amino acids.The amino acid sequence comparison results showed that the leptin receptor amino acid of grass carp shared 35%-86% identity with other species.Phylogenetic tree of leptin receptor generated by Neighbor Joining method suggested that all teleosts cluster together,and the grass carp leptin receptor is clustered closely with zebrafish leptin receptor,goldfish leptin receptor,medaka leptin receptor,fugu rubripes leptin receptor and Atlantic salmon leptin receptor.Then the expression of different tissues including liver,muscle,brain,heart,spleen and mesentery fat tissue were analyzed by real-time fluorogentic quantitative PCR.The results showed that the leptin receptor gene of grass carp expressed in all tissues and highest in spleen（P〈0.05）,lower in heart,brain,muscle and fat tissue,lowest in liver（P〈0.05）.

关 键 词：草鱼 瘦素受体 基因克隆 实时荧光定量PCR 组织表达 

分 类 号：S917.4[农业科学—水产科学] Q78[生物学—分子生物学]