中国水产科学2025，Vol.32Issue(10)：1418-1430,13.DOI:10.12264/JFSC2025-0127
团头鲂phd1基因克隆、分子特征及其在低氧适应中的调控机制
Cloning,molecular characterization,and regulatory mechanism of the phd1 gene in hypoxia adaptation of Megalobrama amblycephala
摘要
Abstract
In this study,we cloned and characterized the prolyl hydroxylase domain protein 1(phd1)gene of Megalobrama amblycephala.The coding sequence measured 1446 bp and encoded 481 amino acids.Bioinformatic analysis indicated that the PHD1 protein had a molecular weight of 52.30 kDa and theoretical isoelectric point of 8.30.It was generally hydrophilic(GRAVY=-0.535),with high proportions of glycine and serine.The secondary structure primarily consisted of random coils(58.00％).Homology modeling of the tertiary structure demonstrated an 82.46％similarity with the template protein(A0A673GA55.1.A).Functional domain analysis identified a conserved 2OG-Fe(Ⅱ)-dependent oxygenase domain,suggesting a potential role for phd1 in hypoxia sensing and regulation.Multiple sequence alignment and phylogenetic analysis indicated that phd1 of M.amblycephala shared the highest similarity(97.72％)with Chanodichthys erythropterus and Culter alburnus,aligning with their taxonomic relationships.Quantitative PCR revealed that phd1 is highly expressed in the liver,heart,and gills(P＜0.05).Under hypoxic stress,phd1 expression in the gills decreased significantly over time,while hif-1αexpression increased(P＜0.05).After 24 h of reoxygenation,both gene expression levels reverted to those observed after 6 h of hypoxia,thereby indicating dynamic regulation of expression in response to oxygen availability.Subcellular localization revealed that the phd1-EGFP fusion protein is localized in the nucleus under both normoxic and hypoxic conditions,thereby suggesting that phd1 may act as a nuclear regulatory factor involved in transcriptional regulation of the HIF signaling pathway.This study elucidates the molecular characteristics of phd1 and its dynamic regulation in response to hypoxia,providing a theoretical basis for understanding adaptive evolution of the HIF pathway in fish.关键词
团头鲂/phd1/生物信息分析/亚细胞定位Key words
Megalobrama amblycephala/phd1/bioinformatics analysis/subcellular localization分类
农业科技引用本文复制引用
FENG Yahui,DING Siyu,CHEN Songlin,LOIC Kemmadzong Foning,ZHENG Guodong,ZOU Shuming..团头鲂phd1基因克隆、分子特征及其在低氧适应中的调控机制[J].中国水产科学,2025,32(10):1418-1430,13.基金项目
国家重点研发计划项目(2022YFD2400600-2) （2022YFD2400600-2）
国家自然科学基金项目(32273100). （32273100）
