水产学报2026，Vol.50Issue(2)：50-64,15.DOI:10.11964/jfc.20250414943
三角帆蚌NQO1基因的克隆、表达及其与Nrf2信号通路的调控关系
Cloning and expression of NQO1 gene and its regulatory relationship with the Nrf2 signaling pathway in Hyriopsis cumingii
摘要
Abstract
This study cloned and functionally characterize the NAD(P)H quinone oxidoreductase 1 gene from Hyriopsis cumingii(HcNQO1),and to elucidated its potential interaction with the H.cumingii transcription factor Nrf2(HcNrf2).The full-length cDNA HcNQO1 was cloned using the rapid amplification of cDNA ends technique.The tissue-specific expression pat-tern of the HcNQO1 mRNA was analyzed by real-time quantitative PCR(RT-qPCR).The promoter sequence of the HcNQO1 was amplified via high-efficiency thermal asymmetric interleaved PCR,and a dual-luciferase reporter assay was used to identify the HcNrf2 binding site.Additionally,protein expression levels of the HcNQO1 in different tissues were assessed using Western blot analysis.The HcNQO1 gene was found to contain an open reading frame of 1 005 bp,encoding a protein of 264 amino acids.Sequence alignment analysis revealed high conservation of HcNQO1 with homologous proteins from other bivalves.RT-qPCR results demonstrated that the HcNQO1 mRNA was expressed in multiple tissues,with significantly higher expression levels observed in the gonads and gills compared to other tissues.A 1 109 bp upstream promoter sequence of HcNQO1 was successfully obtained,and two antioxidant response elements were identified,confirming their essential roles in HcNrf2 binding.Furthermore,polyclonal antibodies against the HcNQO1 protein were generated,and Western blot analysis revealed that the protein expression pattern across tissues differed from that of its mRNA expression.This study successfully cloned and characterized the HcNQO1 gene of H.cumingii and elucidated its interaction with HcNrf2.These findings provide a fundamental theoretical basis for a deeper understanding of the biological function of the NQO1 gene in bivalves and its regu-latory role in oxidative stress responses.关键词
三角帆蚌/Nrf2调控/特异性表达/启动子分析/多克隆抗体/氧化应激Key words
Hyriopsis cumingii/Nrf2 regulation/specific expression/promoter analysis/polyclonal antibodies/oxidative stress分类
农业科技引用本文复制引用
杨清麟,胡绍愚,何雨卓,唐晓琪,余小波,李艳红,吴正理..三角帆蚌NQO1基因的克隆、表达及其与Nrf2信号通路的调控关系[J].水产学报,2026,50(2):50-64,15.基金项目
中国科协青年人才托举工程博士生专项计划(2024) （2024）
重庆市生态渔业产业技术体系(CQMAITS202315) Special Program for Doctoral Students under the Young Talent Support Project of the Chinese Association for Science and Technology(2024) （CQMAITS202315）
Chongqing Ecological Fisheries Industry Technology System(CQMAITS202315) （CQMAITS202315）
