刀鲚PPARγ基因的cDNA克隆及其应激应答
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作者单位:

1. 南京农业大学无锡渔业学院, 江苏 无锡 214081;
2. 中国水产科学研究院淡水渔业研究中心, 农业部淡水渔业和种质资源利用重点实验室, 江苏 无锡 214081

作者简介:

高俊(1992-),男,硕士研究生,从事鱼类规模化繁育及健康养殖技术研究.E-mail:missing_life@163.com

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S965

基金项目:

江苏省成果转化资金专项(BA2015167);江苏省水产三新工程重大专项(D2015-14);中央级公益性科研院所基本科研业务费专项(2017JBFM14).


Molecular cloning and stress response of PPARγ in Coilia nasus
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Affiliation:

1. Wuxi Fisheries College, Nanjing Agricultural University, Wuxi 214081, China;
2. Key Laboratory of Freshwater Fisheries and Germplasm Resources Utilization, Ministry of Agriculture and Rural Affairs;Freshwater Fisheries Research Center, Chinese Academy of Fishery Sciences, Wuxi 214081, China

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    摘要:

    旨在研究刀鲚()过氧化物酶体增殖物激活受体γ(peroxisome proliferator activated receptor,PPARγ)基因的应激调控表达,克隆并获得了刀鲚基因的cDNA全长1951 bp,开放阅读框1470 bp,预测编码489个氨基酸。刀鲚PPARγ包括4个功能结构域,即A/B区,DNA结合区(DNA binding domain,DBD区),铰链区,配体结合区(ligand binding domain,LBD区)。运用荧光定量PCR(real time quantitative PCR,RT-qPCR)检测刀鲚在各组织中均有表达,其中在肝中表达量最高,在脑、肠、心脏、肾、头肾、肌肉中相对高表达,在鳃和脾中微量表达。运输胁迫过程中,在胚胎发育时期各时期均表达,其中在受精卵时期高表达,随后表达量急剧降低,并在此后的时期一直处于较低的表达水平。基因在应激过程中发挥重要作用,也是胚胎发育过程中重要的基因。本研究为刀鲚的人工繁育和应激调控提供了理论基础。

    Abstract:

    is a commercially important fish species that is valued for its nutritive value and is used as a delicacy. It is widely distributed in the Yangtze River, coastal waters of China and Korea, and Ariake Sound in Japan. is highly responsive to stress, which often causes tissue damage, apoptosis, and ultimately death during loading and transport, hampering the development of this industry. Peroxisome proliferator-activated receptors (PPARs) are ligand-dependent transcription factors belonging to the nuclear receptor superfamily. Like other nuclear receptors, PPARs consist of four domains that have distinct functions. The A/B domain, located in conserved N-terminus, is responsible for ligand-independent transcription activity. The C domain, which is the DNA-binding domain (DBD), contains two zinc finger structures. The D domain, the hinge region, connects the DBD with the ligand-binding domain (LBD) and is involved in the conformational changes of PPARs. The E/F domain is an LBD. Transcriptional activation by PPARs requires the presence of PPAR response elements (PPREs) in the promoter of the target gene. PPARs bind PPREs as heterodimers with retinoid X receptor. Three isoforms of PPARs, , have been cloned in various mammals and fish. is involved in many processes such as ontogenesis, lipid metabolism regulation, peroxisome biogenesis, immune functions and inflammation, and oxidative stress. The aim of this study was to investigate the stress response and expression of PPARγ cDNA was obtained, which contained 1951 base pairs with an open reading frame of 1470 base pairs, encoding 489 amino acids. PPARγ has four functional domains, A/B region, DBD, hinge region, and LBD. Real-time quantitative PCR was used to detect the expression of in different tissues and under transport stress. The results showed that was expressed in all tissues, with the highest expression observed in the liver, relatively high expression detected in the brain, intestine, heart, kidney, head kidney, and muscle, and low expression in the gill and spleen. During transport stress, <0.05), peaked at 4 h, and was then significantly down-regulated. High expression of may increase lipid metabolism and relieve inflammation and apoptosis. plays an important role in lipid metabolism and stress responses. During embryonic development, shows the highest expression in the fertilized egg stage, after which its expression is rapidly decreased and then remains at a lower level. Lower expression of may down-regulate lipid metabolism and transportation. This study provides information regarding the theoretical basis of stress regulation.

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高俊,徐钢春,杜富宽,聂志娟,顾若波.刀鲚PPARγ基因的cDNA克隆及其应激应答[J].中国水产科学,2019,26(2):242-250
GAO Jun, XU Gangchun, DU Fukuan, NIE Zhijuan, GU Ruobo. Molecular cloning and stress response of PPARγ in Coilia nasus[J]. Journal of Fishery Sciences of China,2019,26(2):242-250

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  • 在线发布日期: 2019-03-27
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