西伯利亚鲟促黄体激素受体基因的克隆及其生物信息学分析  

Cloning and Bioinformatics Analysis of LHR Gene in Acipenser baerii

作　　者：周洲 孔杰 赵振新 赵飞 吕盛寒 刘霆 ZHOU Zhou;KONG Jie;ZHAO Zhenxin;ZHAO Fei;LYU Shenghan;LIU Ting(Guizhou Fisheries Research Institute,Guiyang,Guizhou 550025;Guizhou Special Aquatic Products Engineering Technology Center,Guiyang,Guizhou 550025,China)

机构地区：[1]贵州省水产研究所,贵州贵阳550025 [2]贵州省特种水产工程技术中心,贵州贵阳550025

出　　处：《贵州农业科学》2025年第1期62-69,共8页Guizhou Agricultural Sciences

基　　金：贵州省基础研究(自然科学)项目“鲟鱼卵子质量蛋白标志物筛选及影响机制研究”(黔科合基础-ZK〔2023〕一般175);国家体系建设项目“特色淡水鱼产业技术体系”(CARS-46)。

摘　　要：【目的】阐明促黄体激素受体(Luteinizing hormone receptor,LHR)的生理功能和LHR基因在西伯利亚鲟性腺发育过程的功能作用,为鲟鱼的生殖调控提供参考。【方法】采用兼并引物扩展及RACE技术,克隆西伯利亚鲟LHR基因cDNA全长,并利用TMHMM、Signal 5.0等在线软件对LHR基因的理化性质、序列同源性等进行生物信息学分析。【结果】西伯利亚鲟LHR基因cDNA序列全长1994 bp,包括开放性阅读框1140 bp,共编码379个氨基酸,相对分子质量为43.02 kD,理论等电点为8.96,有35个正电荷氨基酸残基(Arg+Lys),24个负电荷氨基酸残基(Asp+Glu)。西伯利亚鲟LHR氨基酸序列与小体鲟、大西洋鲟、匙吻鲟和俄罗斯鲟均具有较高的同源性,达92%~98%。LHR蛋白具有7个跨膜结构域,具有G蛋白偶联受体家族蛋白典型保守区域。LHR基因编码的蛋白亲水性平均系数为0.435,属于疏水性氨基酸。二级结构模型显示,LHR蛋白中α-螺旋、无规则卷曲、延伸链以及β-转角的占比分别为41.69%、37.20%、19.53%及1.58%。三级结构模型显示,LHR蛋白主要由α-螺旋和无规则卷曲组成。【结论】从西比利亚鲟卵巢组织成功克隆获得LHR基因cDNA全长序列,LHR氨基酸序列与小体鲟等具有较高的同源性,LHR基因编码的氨基酸序列对应的蛋白质为疏水性氨基酸,存在7个跨膜结构域,具有G蛋白偶联受体家族蛋白典型保守区域,LHR蛋白主要由α-螺旋和无规则卷曲组成。【Objective】The physiological function of luteinizing hormone receptor(LHR)and the function of LHR gene in the gonadal development of Acipenser baerii were studied to provide a reference for the reproductive manipulation of A.baerii.【Method】The full-length cDNA of LHR gene of A.baerii was cloned using degenerate primer extension and RACE techniques,and bioinformatics analysis of the physicochemical properties and sequence homology of LHR gene was carried out by using online software such as TMHMM and Signal 5.0.【Result】The full-length cDNA sequence of LHR gene of A.baerii was 1994 bp,including an open reading frame of 1140 bp,encoding a total of 379 amino acids,with a relative molecular mass of 43.02 kD,a theoretical isoelectric point of 8.96,35 positively-charged amino acid residues(Arg+Lys)and 24 negatively-charged amino acid residues(Asp+Glu).The LHR amino acid sequence of A.baerii had a higher homology with Acipenser ruthenus,Acipenser oxyrinchus,Polyodon spathula and Acipenser gueldenstaedtii,reaching 92%-98%.LHR protein had 7 transmembrane domain with typical conserved regions of G protein-coupled receptor family proteins.The average coefficient of hydrophilicity of the protein encoded by this gene was 0.435,belonging to hydrophobic amino acid.Secondary structure analysis showed that the percentages ofα-helix,random coil,extended strand andβ-turns in the LHR protein were 41.69%,37.20%,19.53%and 1.58%,respectively.The tertiary structure model showed that the LHR protein was mainly composed ofα-helix and random coil.【Conclusion】The full-length cDNA sequence of LHR gene is successfully cloned from A.baerii ovary.LHR amino acid sequence has high homology with A.ruthenus.The protein corresponding to amino acid sequence encoded by LHR gene belongs to hydrophobic amino acid with 7 transmembrane domain and typical conserved regions of G protein-coupled receptor family proteins.LHR protein is mainly composed ofα-helix and random coil.

关 键 词：西伯利亚鲟 促黄体激素 受体 克隆 生物信息学 系统进化树 

分 类 号：S917.4[农业科学—水产科学] TS205[轻工技术与工程—食品科学]