Page 67 - 《水产学报》2026年第2期

2 期 杨清麟，等：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 50 卷

Cloning and expression of NQO1 gene and its regulatory relationship with
the Nrf2 signaling pathway in Hyriopsis cumingii

YANG Qinglin , HU Shaoyu , HE Yuzhuo , TANG Xiaoqi ,
YU Xiaobo , LI Yanhong , WU Zhengli *
(Research Center for Aquatic Biodiversity Conservation in the Upper Reaches of Yangtze River,
College of Fisheries, Southwest University, Chongqing 400715, China)

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.
Key words: Hyriopsis cumingii; Nrf2 regulation; specific expression; promoter analysis; polyclonal antibodies; oxidative stress
Corresponding author: WU Zhengli. E-mail: zh20140202@swu.edu.cn

Funding projects: Special Program for Doctoral Students under the Young Talent Support Project of the Chinese Association
for Science and Technology (2024); Chongqing Ecological Fisheries Industry Technology System (CQMAITS202315)

中国水产学会主办 sponsored by China Society of Fisheries https://www.china-fishery.cn
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