生物学杂志

生物学杂志 ›› 2025, Vol. 42 ›› Issue (4): 81-.doi: 10.3969/j.issn.2095-1736.2025.04.081

• 研究报告 • 上一篇    下一篇

中华绒螯蟹mTORC1抑制性组分PRAS40的cDNA克隆及表达分析

田志环， 焦传珍   

  1. 韶关学院 生物与农业学院， 韶关 512005
  • 出版日期:2025-08-18 发布日期:2025-08-14
  • 通讯作者: 焦传珍，博士，教授，研究方向为生物化学与分子生物学，E-mail:jcz@sgu.edu.cn
  • 作者简介:田志环，博士，教授，研究方向为甲壳动物生长发育，E-mail:tianzhihuan@126.com
  • 基金资助:
    焦传珍，博士，教授，研究方向为生物化学与分子生物学，E-mail:jcz@sgu.edu.cn

Cloning and expression analysis of cDNA encoding the inhibitory component PRAS40 of mTORC1 in Eriocheir sinensis

TIAN Zhihuan, JIAO Chuanzhen   

  1. College of Biology and Agriculture, Shaoguan University, Shaoguan 512005, China
  • Online:2025-08-18 Published:2025-08-14

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摘要： 为探究mTORC1抑制性组分PRAS40在调节甲壳动物生长和代谢中的作用，用RT-PCR技术克隆编码中华绒螯蟹Eriocheir sinensisPRAS40的cDNA序列，并将其编码的蛋白命名为EsPRAS40。EsPRAS40的cDNA编码区序列长1308 bp，编码435个氨基酸。生物信息学分析显示，EsPRAS40 372~432位氨基酸区域含有1个PRAS域、2个分开的长α 螺旋、1个由多个反向β-链和短α-螺旋形成的球状结构域，其余部分（58%）为无规卷曲。系统发育分析显示，EsPRAS40与甲壳动物凡纳对虾（Litopenaeus vannamei）、蚤状溞（Daphnia pulex）和大型蚤（Daphnia magna）的相似序列聚为一支。荧光定量PCR显示，EsPRAS40 mRNA在中华绒螯蟹幼体的Y器、肝胰腺和胃中表达量较高，在眼柄中表达量最低。在饥饿处理的第7天和第14天，EsPRAS40 mRNA在中华绒螯蟹幼蟹的螯足肌肉和肝胰腺组织中表达量显著升高。结果表明，中华绒螯蟹PRAS40可能参与了mTORC1信号通路介导的动物生长和代谢调控。

关键词: 中华绒螯蟹, mTORC1, PRAS40, 克隆, 饥饿

Abstract: To investigate the inhibitory component PRAS40 of mTORC1 in regulating the growth and metabolism on crustaceans, the cDNA sequence encoding PRAS40 inEriocheir sinensiswas cloned using RT-PCR technology, and named the protein as EsPRAS40. The coding sequence of the EsPRAS40 cDNA was 1308 bp long and encoded 435 amino acids. Bioinformatics analysis showed that the EsPRAS40 had a PRAS domain, 2 separate long α-helices, a global structure domain formed by multiple anti-parallel β-chains and short α-helices, and the remaining part (58%) was random coils. Phylogenetic analysis showed that EsPRAS40 clustered with the PRAS40 homologous from the crustaceanLitopenaeus vannamei, Daphnia pulex, andDaphnia magna. Quantitative PCR showed that EsPRAS40 was highly expressed in the Y-organ, hepatopancreas, and stomach ofE. sinensislarvae, with the lowest expression in the eye stalk. The expression level of EsPRAS40 mRNA was significantly increased in the claws and hepatopancreas of larvalE. sinensisafter 7 days and 14 days of fasting. The results in this study indicated that the putative mTORC1 inhibitory component PRAS40 involved in regulating the growth and metabolism controlled by mTOR signaling pathway inE. sinensis.

Key words: Eriocheir sinensis, mTORC1, PRAS40, clone, fasting

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