| 中华鳖GnRH 1基因cDNA克隆及表达特征 |
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| 引用本文： | 申慧,张英萍,王利华,沙航,王咏星,邹桂伟,梁宏伟.中华鳖GnRH 1基因cDNA克隆及表达特征[J].淡水渔业,2019(4):16-21. |
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| 作者姓名： | 申慧 张英萍 王利华 沙航 王咏星 邹桂伟 梁宏伟 |
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| 作者单位： | 新疆大学生命科学与技术学院;中国水产科学研究院长江水产研究所;上海海洋大学水产与生命学院 |
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| 基金项目： | 安徽省科技重大专项(16030701065);水产种质资源平台项目(2017DKA30470) |
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| 摘 要： | 为了解GnRH基因在中华鳖(Pelodiscus sinensis)性腺和胚胎发育过程中的表达特征,采用cDNA末端快速扩增(RACE)技术从中华鳖全脑中获得与生长生殖调控密切相关的GnRH1基因全长cDNA,并运用实时荧光定量PCR(qRT-PCR)技术检测GnRH1在成鳖不同组织和胚胎发育时期的表达水平。结果显示:中华鳖GnRH1基因cDNA全长546 bp,其中5′非编码区(5′UTR)99 bp,3′非编码区(3′UTR)168 bp,开放阅读框(ORF)279 bp,编码92个氨基酸,分子质量为10.23 ku,理论等电点pI为5.65,具有N端信号肽(1~23 aa)、核心十肽区域(24~33 aa)、断裂位点GKR(34~36 aa)及相关肽区域(37~92 aa),符合GnRH蛋白典型结构特征。系统进化树结果显示,中华鳖GnRH1基因和绿海龟(Chelonia mydas)、墨西哥箱龟(Terrapene carolina mexicana)及西部锦龟(Chrysemys picta bellii)GnRH1基因聚为一支。qRT-PCR结果表明,GnRH1基因在中华鳖雌雄个体的8个组织中均有表达,在脑和性腺组织中高表达,且具有性别差异,雄性中华鳖中的表达显著高于雌性(P<0.05);在10个胚胎发育时期均表达,且随发育时间的后移,表达量显著增加,在第16期达到峰值。GnRH1基因可能在中华鳖生长及性腺分化中具有重要作用。
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| 关 键 词： | 中华鳖(Pelodiscus sinensis) 促性腺激素释放激素 基因克隆 基因表达 |
Molecular cloning and expression analysis of the GnRH 1 cDNA in Pelodiscus sinensis |
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| Institution: | (College of Life Science and Technology,Xinjiang University,Urumqi 830046,China;Yangtze River Fisheries Research Institute,Chinese Academy of Fishery Sciences,Wuhan 430223,China;College of Fisheries and life Science,Shanghai Ocean University,Shanghai 201306,China) |
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| Abstract: | Gonadotropin releasing hormone ( GnRH )is an important regulator of reproductive development in animals.The GnRH1 cDNA were cloned by rapid amplification cDNA ends ( RACE)and the expression characteristics of the gene by Quantitative real-time PCR ( qRT-PCR)were analyzed to analyze features and functions of the GnRH1 gene in the gonad developmen of Pelodiscus sinensis.The results showed that the full length of GnRH1 cDNA sequence consisted of546 bp,which contained 99 bp5′ untranslated region ( UTR),169 bp3′ UTR,and an open reading frame of279 bp,encoded a protein with 92 amino acids. GnRH1 included a putative signal peptide (1~23 aa),a core decapeptide region (24~33 aa),a cleavage site GKR (34~36 aa)and GnRH associated peptide ( GAP)(37~92 aa).Phylogenetic analysis showed that the GnRH1 gene was clustered in the same subgroup with the reptile.qRT-PCR results indicated that the GnRH1 gene were expressed widely in all 8 tissues in adults,the highest expression level was detected in the brain and gonad.The expression pattern exhibited sexual dimorphism.The expression level of GnRH1 were significantly higher in male ( P <0.05).The highest expression was found in the sixteenth period of embryo development.Results indicated that the GnRH1 gene might play an important role in the growth and gonadal development of P.sinensis . |
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| Keywords: | Pelodiscus sinensis gonadotropin releasing hormone1 cloning expression pattern |
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