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2025年8月6日 星期三
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半滑舌鳎甲状腺激素受体β基因(TRβ)的克隆与表达分析
刘琨1,2,陈松林2,张丽燕2,董忠典2,李海龙2,刘万军2
1. 上海海洋大学水产与生命学院
2. 中国水产科学研究院黄海水产研究所
Molecular Cloning and Expression of Thyroid Hormone Receptors β Gene(TRβ) from Half-smooth Tongue Sole (Cynoglossus semilaevis)
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摘要 甲状腺激素在鲽形目仔鱼的变态过程中起着重要的作用,而甲状腺激素的生物效应是由甲状腺激素受体(thyroid hormone receptors, TRs)介导的。本研究利用RACE技术克隆了半滑舌鳎(Cynoglossus semilaevis)TRβ基因的全长,并对其功能进行了生物信息学分析。结果显示,TRβ 基因(GenBank登录号: No. KJ499433) cDNA全长为2 345 bp,开放读框(open reading frame, ORF)长1 188 bp,编码396个氨基酸,5'-UTR 和3'-UTR 分别长454和703 bp。通过氨基酸序列比对发现,TRβ 氨基酸序列铰链区有一段由9个氨基酸残基组成的插入序列,推测这段序列可能为硬骨鱼类TRβ 基因所特有的。实时荧光定量PCR技术分析结果显示,TRβ 基因在各组织中均有表达,且在肝组织中的表达量显著高于其他组织(P<0.05);因此推测TRβ 基因在半滑舌鳎成鱼各组织中具有多样性的功能,而且其在肝脏的新陈代谢中起到重要的调节作用。在半滑舌鳎仔鱼变态时,TRβ基因的表达量随变态开始而迅速增加,在变态高峰期到达最高。药物处理实验结果显示,正常变态和提前变态的仔鱼TRβ 基因的表达水平均随变态开始而迅速上调,而变态被抑制的仔鱼中TRβ 基因的表达量则未出现显著变化,表明TRβ 是调控半滑舌鳎仔鱼变态过程的重要基因,在半滑舌鳎生长发育早期阶段起较关键作用。本研究为深入解析半滑舌鳎变态的分子机制提供了理论依据和参考。
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刘琨
陈松林
张丽燕
董忠典
李海龙
刘万军
关键词 半滑舌鳎甲状腺激素受体β变态基因克隆表达    
Abstract:Thyroid hormone plays an important role in the metamorphosis of flatfish and its biological effects are mediated by thyroid hormone receptors (TRs). In this study, the full length of TRβ had been sequenced in Cynoglossus semilaevis and its biological activities were analyzed. The nucleotide sequence of TRβ was 2 345 bp, including 5' terminal UTR of 454 bp, 3' terminal UTR of 703 bp and complete open reading frame of 1 188 bp which encoded 396 amino acid residues. Compared to the amino acid sequences of TRs in different species, we found a insertion of 9 amino acids in the hinge region which was unique to TRβ of teleosts. We found that the expression level of TRβ in liver was significantly higher than that in other tissues using Real-time PCR. We speculated that TRβ had diverse functions in tissues of adult Cynoglossus semilaevis, especially might play an important role in liver metabolism. We also found that TRβ gene transcript increased rapidly in the beginning of metamorphosis, reached peak in metamorphic climax. Moreover, larvae were exposed to the thyroid hormone(T4) and goitrogen thiourea(TU) and the expression levels of TRβ were detected. The expression levels of TRβ in control and T4-treated group rapidly increased as metamorphosis began. On the contrary, the expression levels of TRβ in TU-treated group still remained in a low level. In conclusion, TRβ gene played an important role in the metamorphosis of Cynoglossus semilaevis. The research provides a theoretical basis and reference for molecular mechanism research of metamorphosis of Cynoglossus semilaevis.
Key wordsCynoglossus semilaevis    Thyroid hormone receptor β    Metamorphosis    Gene cloning    Expression level
收稿日期: 2014-03-03     
通讯作者: 陈松林   
引用本文:   
刘琨1,2,陈松林2,张丽燕2,董忠典2,李海龙2,刘万军2. 半滑舌鳎甲状腺激素受体β基因(TRβ)的克隆与表达分析[J]. , 2014, 22(9): 1157-1165.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2014/V22/I9/1157
 
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