暗纹东方鲀抗苗勒氏管激素Ⅱ型受体基因的克隆、生物信息学及表达分析  

Molecular cloning, bioinformatics and expression analysis of anti-Müllerian hormone type Ⅱ receptor gene in Takifugu obscures

作　　者：高莹莹 胡鹏 刘新富[1] 黄滨[1] 刘滨[1] GAO Ying-ying;HU Peng;LIU Xin-fu;HUANG Bin;LIU Bin(Yellow Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences,Qingdao Shandong 266071,China;College of Fishery and Life Sciences,Shanghai Ocean University,Shanghai 201306,China;Neijiang Normal University,Neijiang Sichuan 641100,China)

机构地区：[1]中国水产科学研究院黄海水产研究所,山东青岛266071 [2]上海海洋大学水产与生命学院,上海201306 [3]内江师范学院,四川内江641100

出　　处：《海洋渔业》2019年第5期555-566,共12页Marine Fisheries

基　　金：国家重点研发计划(2017YFB0404001);国家海水鱼产业技术体系(CARS-47-G24)

摘　　要：为探究暗纹东方鲀(Takifugu obscures)抗苗勒氏管激素Ⅱ型受体(anti-Müllerian hormone receptorⅡ,Amhr2)基因的序列和结构信息,初步研究amhr2基因在性腺发育和性别分化过程中的作用,实验通过设计简并引物扩增及RACE技术,克隆出暗纹东方鲀amhr2完整的CDS区,分析其相应的生物信息学特征及其在不同组织和不同发育期的mRNA表达水平。结果表明:amhr2序列cDNA全长为1 868 bp(NCBI登录号:MH218814),编码513个氨基酸,与红鳍东方鲀(Takifugu rubripes)同源性最高,达99%。氨基酸多重序列比对结果显示,Amhr2氨基酸序列中的近C-末端区域序列较为保守;通过构建系统进化树发现,暗纹东方鲀Amhr2与红鳍东方鲀亲缘关系最近。荧光定量PCR分析显示,暗纹东方鲀amhr2仅在性腺中表达,且卵巢中的表达量明显高于精巢;并且在性腺早期发育过程中,amhr2在精巢和卵巢中的表达量均呈现出先降低再迅速升高的表达趋势。研究结果表明,amhr2在暗纹东方鲀中的表达具有组织特异性,且在性别分化和性别维持等过程中发挥重要的作用。In order to investigate the sequence and structural information of the anti-Müllerian hormone receptor Ⅱ(Amhr2) gene in Takifugu obscurus, preliminarily study the role of amhr2 gene in gonad development and sex differentiation, we successfully cloned the intact CDS region of the amhr2 gene in T.obscurus through the design of degenerate primer amplification and RACE technology, subsequently its corresponding bioinformatics characteristics and the level of mRNA expression in different tissues and developmental stages were also analyzed. The results showed that the full-length sequence of amhr2 gene in T.obscurus was 1 868 bp(NCBI accession number: MH218814), encoding 513 amino acids, and sharing the highest homology(99%) with the gene of Takifugu rubripes. The results of multiple amino acid sequence alignment showed that the part of the near C-terminal region in the Amhr2 amino acid sequence was more conservative. Furthermore, the phylogenetic tree analysis of the Amhr2 amino acid sequence revealed the closest relationship between T.obscures and T.rubripes. The fluorescence quantitative PCR analysis showed that amhr2 gene in T.obscurus was only expressed in the gonad, and the expression level in ovary was significantly higher than that in testis. During the early development of gonad, the expression of amhr2 in testis and ovary showed a trend of decreasing first and then rising rapidly. The results showed that the expression of amhr2 gene was tissue-specific and played an important role during sex differentiation and sex maintenance.

关 键 词：暗纹东方鲀 抗苗勒氏管激素Ⅱ型受体 克隆 生物信息学 组织表达 发育期 

分 类 号：S917.4[农业科学—水产科学] Q786[生物学—分子生物学]