大刺鳅mstn基因克隆及表达分析Molecular Cloning and Expression Profiles of Myostatin Gene in Mastacembelus armatus
钟东明,赖星星,张明清,舒琥
摘要(Abstract):
【目的】了解大刺鳅(Mastacembelus armatus)肌肉生长抑制素(myostatin)基因mstn的序列及其在生长发育中的调控功能。【方法】利用RACE方法克隆大刺鳅mstn全长cDNA序列,采用荧光定量PCR分析其在不同组织和胚胎发育阶段的表达情况。【结果与结论】大刺鳅mstn cDNA序列全长2 690 bp,包含200 bp 5′非编码区、1 359 bp 3′非编码区和1 131 bp开放阅读框,编码376个氨基酸。前22个氨基酸残基为信号肽,第37~254位氨基酸残基为TGF-β前肽域,第282~376位是TGF-β结构域,具蛋白酶水解位点RARR和C端生物活性区的9个保守半胱氨酸残基。系统发育分析表明,大刺鳅MSTN氨基酸序列与合鳃目、鲈形目鱼类同源性较高,与哺乳动物、鸟类、爬行类、两栖类同源性较低。实时定量PCR分析表明,大刺鳅mstn基因在各组织中均有表达,肌肉中表达量最高,眼、脑次之,鳃表达量最低;大刺鳅胚胎发育的各阶段mstn基因均有表达,囊胚期表达量最高,其次为多细胞期,出膜期表达量最低。
关键词(KeyWords): 大刺鳅;mstn;基因;克隆;表达
基金项目(Foundation): 中国-东盟海上合作基金项目(2018.01—2020.12);; 国家公益性行业(农业)重大专项(201303048);; 广东省海洋渔业科技攻关与研发项目(A201601A05;2017A0007)
作者(Author): 钟东明,赖星星,张明清,舒琥
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