大口黑鲈gyk和foxq1基因克隆及营养调控分析  

Cloning of gyk and foxq1 Gene and Analysis of Nutritional Regulation in Micropterus salmoides

作　　者：宋含茹 雷彩霞 阎瀚威 朱聪聪 宋红梅[2] 杜金星 朱涛 采克俊[1] 李胜杰[2] SONG Hanru;LEI Caixia;YAN Hanwei;ZHU Congcong;SONG Hongmei;DU Jinxing;ZHU Tao;CAI Kejun;LI Shengjie(School of Life Sciences,Huzhou Normal University,Huzhou,313000;Key Laboratory of Ministry of Agriculture and Rural Affairs for Tropical and Subtropical Fishery Resource Application and Cultivation,Guangdong Provincial Key Laboratory of Aquatic Animal Immunology and Sustainable Aquaculture,Pearl River Fisheries Research Institute,Chinese Academy of Fishery Sciences,Guangzhou,510380;College of Fisheries and Life Science,Shanghai Ocean University,Shanghai,201306)

机构地区：[1]湖州师范学院生命科学学院,湖州313000 [2]中国水产科学研究院珠江水产研究所,农业农村部热带亚热带水产资源利用与养殖重点实验室,广东省水产动物免疫与绿色养殖重点实验室,广州510380 [3]上海海洋大学水产与生命学院,上海201306

出　　处：《基因组学与应用生物学》2025年第2期170-181,共12页Genomics and Applied Biology

基　　金：广东省基础与应用基础研究基金项目(2023A1515010215);中国水产科学研究院中央级公益性科研院所基本科研业务费专项(2024SJRC3);2024年省级乡村振兴战略专项资金种业振兴项目(2024-SPY-00-003);广州市科技计划项目(2023A04J0092)共同资助。

摘　　要：哺乳动物甘油激酶(glycerokinase,GYK)和叉头框Q1(forkhead box Q1,FOXQ1)在调节机体糖代谢中发挥重要作用。为探索gyk和foxq1基因在大口黑鲈(Micropterus salmoides)糖稳态调节中的作用,本研究克隆了gyk基因5′端侧翼序列2056 bp,完整编码区序列1686 bp,以及foxq1基因完整编码区序列843 bp,并进行了营养调控分析。多重序列比对和系统进化树分析显示,不同物种间gyk基因具有高度的同源性,而foxq1仅在结构域中与其他物种具有较高的同源性。通过生物信息学预测分析发现gyk基因5′端含有FOXQ1、TFAP2A和ARNT等多个转录因子结合位点;gyk基因含有2个FGGY结构域;foxq1基因含1个Fox结构域。实时定量PCR(real-time quantitative PCR,RT-qPCR)结果表明,gyk在大口黑鲈肌肉组织中的表达量最高,其次为肝脏、脂肪和心脏,在脾和肾中表达量最低;foxq1在肌肉和肝脏中表达量最高,在鳃、肾、胃和脾中表达量最低。饱食投喂大口黑鲈24 h内,gyk和foxq1基因在肝脏和肌肉组织中的表达量和血糖含量均呈先上升后下降的变化趋势,且在摄食后12 h时gyk和foxq1基因表达量和血糖含量达到最高。大口黑鲈分别摄食高淀粉水平和低淀粉水平的两种饲料,高淀粉组中肝脏和肌肉组织的gyk、foxq1的表达量均显著高于低淀粉组的。研究结果表明gyk和foxq1基因可能参与大口黑鲈糖稳态调节,且FOXQ1可能作为转录因子参与调节gyk的转录活性,进而调控大口黑鲈糖代谢。研究结果为深入解析gyk和foxq1基因功能及其在大口黑鲈营养育种中的应用提供基础数据和理论依据。Mammalian glycerokinase(GYK)and forkhead box Q1(FOXQ1)play important roles in the regulation of organismal glucose metabolism.In order to explore the roles of gyk and foxq1 genes in the regulation of glucose homeostasis in largemouth bass(Micropterus salmoides),we cloned 2056 bp of flanking sequence at the 5′end of gyk gene and 1686 bp of complete coding region sequence;and the complete coding region sequence of foxq1 gene was 843 bp.In addition,a nutritional regulation analysis was carried out.Multiple sequence comparison and phylogenetic tree analysis showed that gyk gene was highly homologous among different species,while foxq1 was only highly homologous to other species in its structural domains.Bioinformatics prediction analysis revealed that the 5′end of gyk gene contained multiple transcription factor binding sites such as FOXQ1,TFAP2A and ARNT;gyk gene contained two FGGY structural domains;foxq1 gene contained one Fox structural domain.The results of real-time quantitative PCR(RT-qPCR)showed that the expression of gyk was the highest in muscle tissue of Micropterus salmoides,followed by liver,fat and heart,and the lowest in spleen and kidney;foxq1 had the highest expression in muscle and liver,and the lowest expression in gill,kidney,stomach and spleen.The expression of gyk and foxq1 genes in liver and muscle tissues and blood sugar content of Micropterus salmoides showed a tendency of increasing and then decreasing within 24 h of feeding,and the expression of gyk and foxq1 genes and blood sugar content reached the highest at 12 h of feeding.When Micropterus salmoides were two types of diets,one with a high-starch level and the other with a low-starch level respectively,the expression of gyk and foxq1 in liver and muscle tissues were significantly higher in the high starch group than in the low starch group.The results indicated that gyk and foxq1 genes may be involved in the regulation of sugar homeostasis in Micropterus salmoides,and that FOXQ1 may act as a transcription factor to regulate the trans

关 键 词：大口黑鲈 营养调控 克隆 表达分析 

分 类 号：S917.4[农业科学—水产科学]