梭鲈ho1基因的克隆及其低氧胁迫下表达分析  

Characterization and expression analysis of ho1 from Sander lucioperca under acute hypoxia stress

作　　者：吉宇丹 孙志鹏[1] 吕伟华[1] 鲁翠云[1,2] 曹顶臣[1] 刘天奇 周佳 郑先虎[1,2] JI Yudan;SUN Zhipeng;LYU Weihua;LU Cuiyun;CAO Dingchen;LIU Tianqi;ZHOU Jia;ZHENG Xianhu(Heilongjiang River Fisheries Research Institute,Chinese Academy of Fishery Sciences/Key laboratory of Freshwater Aquatic Biotechnology and Breeding,Ministry of Agriculture and Rural Affairs,Harbin 150070,China;Shanghai Ocean University/Key Laboratory of Freshwater Aquatic Germplasm Resources,Ministry of Agriculture and Rural Affairs,Shanghai 201306,China)

机构地区：[1]中国水产科学研究院黑龙江水产研究所/农业农村部淡水水产生物技术与遗传育种重点实验室,黑龙江哈尔滨150070 [2]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

出　　处：《南方水产科学》2023年第2期98-106,共9页South China Fisheries Science

基　　金：中国水产科学研究院黑龙江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(HSY202009Q);国家重点研发计划项目(2019YFD0900405);中央引导地方科技发展专项(ZY21C03);国家淡水水产种质资源库项目(FGRC18537)。

摘　　要：梭鲈(Sander lucioperca)对低氧极敏感,在集约化养殖和苗种运输过程中易发生低氧应激和死亡现象。为探究血红素加氧酶1(Heme oxygenase 1,HO1)在梭鲈响应低氧过程中的调节作用,通过RACE(Rapid amplification of cDNA ends)技术克隆了梭鲈ho1基因,其cDNA全长为1256 bp,包含840 bp的开放阅读框(Open reading frame,ORF)、162 bp的5'非编码区(Untranslated region,5'-UTR)和254 bp的3'-UTR,编码279个氨基酸。多重序列比对显示,梭鲈HO1与翘嘴鳜(Siniperca chuatsi)、舌齿鲈(Dicentrarchus labrax)和大口黑鲈(Micropterus salmoides)的氨基酸序列相似性分别为91.84%、88.69%和88.11%。实时荧光定量结果显示,ho1基因在所有检测组织中均有表达,其中在脑组织中高表达,其次是肾、肝、鳃等组织。低氧刺激前3 h,梭鲈ho1主要在皮肤、鳃中响应;低氧胁迫3 h后,ho1主要在心、肝、肾中发挥转录调控作用。复氧12 h,除肝脏外,梭鲈其他组织ho1的相对表达量均可恢复正常,低氧刺激对肝组织ho1的表达产生了较大影响。研究表明,ho1基因参与梭鲈响应低氧的分子调节机制并在其中发挥着重要的生物学功能,可为深入了解梭鲈低氧胁迫遗传机制提供理论参考。Sander lucioperca is extremely sensitive to hypoxia,and is prone to hypoxia stress and death during intensive breeding and seedling transportation.In order to investigate the regulating effect of heme oxygenase 1(HO1)in the response to hypoxia of S.lucioperca,we cloned the full-length cDNA sequence of ho1 gene by RACE(Rapid amplification of cDNA ends)technology.The results indicate that the cDNA length was 1256 bp(840 bp ORF,162 bp 5'-UTR and 254 bp 3'-UTR),encoding 279 amino acids.The multiple sequence alignment shows that the similarity of HO1 with Siniperca chuatsi,Dicentrarchus labrax and Micropterus salmoides was 91.84%,88.69%and 88.11%,respectively.Real-time quantitative PCR discloses that ho1 was expressed in all the tested tissues,with the highest concentration in the brain,followed by the kidney,liver and gills.During the first 3-hour hypoxic stimulation of Pikeperch,ho1 primarily responded in the skin and gills,but mainly played transcriptional regulatory roles in the heart,liver and kidney after 3 h of hypoxic stress.At 12th hour of reoxygenation,the expression levels of ho1 in all the tissues except the liver returned to a normal level,and hypoxia stress had an enormous effect on the expression of ho1 in the liver.The study reveals that ho1 gene is involved in the molecular regulation mechanisms of S.lucioperca in response to hypoxia and plays an important biological role,which provides theoretical references for understanding the genetic mechanism of hypoxic stress.

关 键 词：梭鲈 ho1 基因克隆 低氧胁迫 基因表达 

分 类 号：S965.1[农业科学—水产养殖]