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中华绒螯蟹C/EBPβ基因的克隆及表达

杨志刚 孙小万 陈阿琴 李腾 王爱民 成永旭

中国水产科学2026,Vol.33Issue(4):1-12,12.
中国水产科学2026,Vol.33Issue(4):1-12,12.DOI:10.12264/JFSC2025-0342

中华绒螯蟹C/EBPβ基因的克隆及表达

Cloning and expression analysis of the C/EBPβ gene in Eriocheir sinensis

杨志刚 1孙小万 1陈阿琴 1李腾 1王爱民 2成永旭3

作者信息

  • 1. 上海海洋大学,农业农村部淡水水产种质资源重点实验室,上海 201306||上海海洋大学,农业农村部鱼类营养与环境生态研究中心,上海 201306
  • 2. 盐城工学院海洋与生物工程学院,江苏 盐城 224000
  • 3. 宁波大学海洋学院,浙江 宁波 315832
  • 折叠

摘要

Abstract

We investigated the molecular regulatory role of CCAAT/enhancer-binding protein β(C/EBPβ),an important bZIP transcription factor,in Eriocheir sinensis lipid metabolism.The full-length cDNA of C/EBPβ was obtained by reverse transcription PCR combined with rapid amplification of cDNA ends using hepatopancreas cDNA as the template.Bioinformatic analyses were performed to characterize sequence features and predict structural and physicochemical properties of the protein.The tissue expression profile of C/EBPβ was examined using quantitative PCR.Furthermore,functional verification was conducted via injections of the C/EBPβ agonist Rosiglitazone,inhibitor Withaferin A,and siRNA-mediated gene silencing to evaluate regulatory effects of C/EBPβ on the expression of lipid metabolism–related genes,including Fad6,Fad9,Elovl4,Elovl6,and Elovl7.The full-length cDNA of C/EBPβ was 1637 bp,containing an 879-bp open reading frame encoding a 293-amino-acid protein,a 227-bp 5′-UTR,and a 531-bp 3′-UTR.Sequence analysis revealed a conserved bZIP domain,confirming its identity as a member of the C/EBP family.Homology analysis revealed high similarity of E.sinensis C/EBPβ with Portunus trituberculatus(78.23%)and Chionoecetes opilio(79.93%).Phylogenetic analysis revealed that E.sinensis C/EBPβ clustered closely with that of other crab species.Tissue distribution analysis revealed significantly higher C/EBPβ expression in the hepatopancreas and gills than in other tissues(P<0.05).Functional assays demonstrated that Rosiglitazone injection significantly upregulated Fad6,Fad9,Elovl6,and Elovl7 expression,whereas Withaferin A and siRNA interference markedly downregulated their transcription levels(P<0.05).Elovl4 expression exhibited no significant response to any treatment.In this study,we cloned the full-length C/EBPβ gene from E.sinensis and confirmed that C/EBPβ could promote LC-PUFA biosynthesis by regulating transcription of desaturase and elongase genes in the hepatopancreas.These results provide new insights into the biological functions of C/EBPβ in crustaceans and offer a theoretical basis for elucidating the molecular mechanisms underlying lipid metabolism regulation in E.sinensis.

关键词

中华绒螯蟹/C/EBPβ/基因克隆/脂质代谢/LC-PUFA

Key words

Eriocheir sinensis/C/EBPβ/gene cloning/lipid metabolism/LC-PUFA

分类

农业科技

引用本文复制引用

杨志刚,孙小万,陈阿琴,李腾,王爱民,成永旭..中华绒螯蟹C/EBPβ基因的克隆及表达[J].中国水产科学,2026,33(4):1-12,12.

基金项目

国家自然科学基金项目(32273154) (32273154)

现代农业产业技术体系项目(CARS-48). (CARS-48)

中国水产科学

1005-8737

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