中华绒螯蟹Rheb编码序列的克隆及表达分析
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韶关学院生物与农业学院, 广东 韶关 512005

作者简介:

田志环(1971-),博士,教授,研究方向为甲壳动物生长发育.E-mail:tianzhihuan@126.com

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S917

基金项目:

广东省驻镇帮扶农村科技特派员项目(KTP20210401); 韶关学院重点科研项目(432-99000440); 韶关学院博士引进项目(432-99000622)


Cloning and expression analysis of sequence encoding Rheb in Eriocheir sinensis
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College of Biology and Agriculture, Shaoguan University, Shaoguan 512005 , China

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    摘要:

    Rheb (Ras homologue enriched in brain, 脑中富集的 Ras 同源物)是 mTOR (mammalian or mechanistic target of rapamycin, 哺乳动物雷帕霉素靶蛋白)信号通路上游重要的调控因子。为探究 Rheb 通过 mTOR 信号通路调控中华绒螯蟹(Eriocheir sinensis)生长发育和营养代谢的作用, 本研究采用 RT-PCR 克隆了中华绒螯蟹 Rheb (EsRheb)的 cDNA 编码序列, 并用生物信息学方法分析了其序列和结构特征, 用反转录荧光定量 PCR 检测了 EsRheb mRNA 的组织分布及饥饿和摘除眼柄对 EsRheb 转录水平的影响。结果显示 EsRheb 含 182 个氨基酸, 其氨基酸序列、保守结构域、功能位点与其他不同生物的 Rheb 非常相似, 三维结构和人 Rheb 高度一致。EsRheb 的 mRNA 在中华绒螯蟹肝胰腺、胃、心肌、螯足肌肉和 Y 器中表达较高, 在眼柄中表达最低。饥饿处理显著降低螯足肌肉 EsRheb 的转录表达水平, 对肝胰腺 EsRheb 的转录表达影响不显著; 摘除眼柄显著提高 EsRheb 在 Y 器和螯足肌肉中的转录表达水平。该结果为进一步阐明 Rheb 参与中华绒螯蟹 mTOR 介导的生长发育和营养代谢调控奠定了基础。

    Abstract:

    Ras homologue enriched in the brain (Rheb) is an important positive regulator on the upstream of the mammalian mechanistic target of rapamycin (mTOR) signaling pathway. The cDNA sequence encoding Rheb in Eriocheir sinensis (EsRheb) was cloned using RT-PCR in order to investigate the role of this gene in the regulation of growth, development, and nutrient metabolism of Eriocheir sinensis via the mTOR signaling pathway. The sequence and structural characteristics of EsRheb were analyzed using bioinformatics. The tissue distribution of EsRheb mRNA and the effects of animal starvation and eyestalk ablation on EsRheb transcription were detected using qRT-PCR. The results showed that EsRheb cDNA encoded 182 amino acids; its amino acid sequence, conserved domain, and functional sites were highly similar to the Rheb of a wide range of species; and its three-dimensional structure was highly consistent with that of human Rheb. EsRheb mRNA expression was high in the hepatopancreas, stomach, myocardium, skeletal muscles, and Y organ, but was much lower in the eye stalk. Starvation treatment of animals significantly decreased the transcriptional expression of EsRheb in the skeletal muscle but had no significant effect on its expression in the hepatopancreas. The removal of the eye stalk significantly increased the transcriptional expression of EsRheb in the Y organs and skeletal muscles. This study provides a basis for elucidating the function of EsRheb in regulating the mTOR-mediated growth and metabolism of E. sinensis.

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引用本文

田志环,焦传珍.中华绒螯蟹Rheb编码序列的克隆及表达分析[J].中国水产科学,2024,31(12):1432-1440
TIAN Zhihuan, JIAO Chuanzhen. Cloning and expression analysis of sequence encoding Rheb in Eriocheir sinensis[J]. Journal of Fishery Sciences of China,2024,31(12):1432-1440

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  • 收稿日期:2024-06-24
  • 最后修改日期:2024-09-10
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  • 在线发布日期: 2025-02-11
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