摘要
凋亡抑制因子1 (tumor protein p53 regulated inhibitor of apoptosis 1, TRIAP1)可以通过抑制细胞凋亡参与生物体内应激反应。为探究TRIAP1 基因及调控其表达的miRNAs 在斑节对虾(Penaeus monodon)应对哈氏弧菌(Vibrio harveyi)刺激时的表达调控情况,该研究通过RACE 技术获得了斑节对虾TRIAP1 基因(PmTRIAP1)cDNA 全长序列,并利用实时荧光定量PCR (qRT-PCR)技术,对PmTRIAP1 在斑节对虾不同组织中的表达分布模式,以及哈氏弧菌刺激下PmTRIAP1 与相关miRNAs 的表达关联情况进行了探究。结果显示,PmTRIAP1cDNA 全长2 522 bp,包括11 bp 的5'非编码区(UTR)、2 289 bp 的3'UTR 和222 bp 的开放阅读框(ORF),共编码73 个氨基酸。PmTRIAP1 基因在各组织中均有表达,其中在血细胞中表达量最高;哈氏弧菌刺激下,PmTRIAP1表达量显著降低,而Pm-miR-145-3p 和Pm-miR-454-3p 表达量显著升高,表明PmTRIAP1 和Pm-miR-145-3p、Pm-miR-454-3p 的表达量呈负相关关系;双荧光素酶报告实验结果显示,Pm-miR-145-3p 和Pm-miR-454-3p 可通过结合PmTRIAP1 3'UTR 降低荧光素酶活性,表明Pm-miR-145-3p 和Pm-miR-454-3p 均可靶向调控PmTRIAP1 的表达。
TRIAP1 can participate in vivo responses by inhibiting apoptosis. In order to analyze the expression regulation of TRIAP1 and miRNAs that regulate TRIAP1's expression under Vibrio harveyi challenge in Penaeus monodon, we obtained the P. monodon TRIAP1 gene (PmTRIAP1) cDNA by RACE, then detected the tissue distribution of PmTRIAP1 by quantitative real-time PCR (qRT-PCR), and investigated the expression association between PmTRIAP1 and its related miRNAs under V. harveyi challenge. The results show that the full-length PmTRIAP1 cDNA sequence was 2 522 bp, containing an 11 bp 5' non-coding region (UTR), a 2 289 bp 3'-UTR and a 222 bp open reading frame encoding 73 amino acids. The qRT-PCR analysis shows that PmTRIAP1 was ubiquitously expressed in all the tested tissues and the highest expression level was obtained in hemocyte. The expression of PmTRIAP1 decreased significantly;however, the expressions of Pm-miR-145-3p and Pm-miR-454-3p increased significantly under V. harveyi challenge, which suggests that the expression levels of PmTRIAP1 and Pm-miR-145-3p, Pm-miR-454-3p were negatively correlated. Dual luciferase reporter assay shows that Pm-miR-145-3p and Pm-miR-454-3p can bind PmTRIAP1 3'UTR to reduce luciferase activity. It is suggested that both Pm-miR-145-3p and Pm-miR-454-3p can target regulate the expression of PmTRIAP1.
作者
刘恩瑞
赵超
王鹏飞
范嗣刚
闫路路
邱丽华
LIU Enrui;ZHAO Chao;WANG Pengfei;FAN Sigang;YAN Lulu;QIU Lihua(Key Laboratory of South China Sea Fishery Resources Exploitation & Utilization,Ministry of Agriculture and Rural Affairs,South China Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences,Guangzhou 510300,China;National Demonstration Center for Experimental Fisheries Science Education,Shanghai Ocean University,Shanghai 201306,China;Key Laboratory of Aquatic Genomics,Ministry of Agriculture and Rural Affairs,Beijing 100141,China)
出处
《南方水产科学》
CAS
CSCD
北大核心
2019年第4期88-98,共11页
South China Fisheries Science
基金
国家重点研发计划项目(2018YFD0900303)
中国水产科学研究院南海水产研究所中央级公益性科研院所基本科研业务费专项资金资助(2019TS11)
海南自然科学基金创新研究团队项目(2017CXTD021)
海南省重点研发计划项目(ZDYF2018163)
广东省海洋渔业科技与产业发展专项(A201701A04)
