黄颡鱼脂代谢关键基因PPARα的启动子克隆及功能初探
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- 英文论文题名：Molecular cloning and functional analysis of PPARα promoter in yellow catfish Pelteobagrus fulvidraco
- 论文作者：有文静 ; 范耀方 ; 谭肖英 ; 徐异桓 ; 吴坤 ; 魏川川
- 英文论文作者：YOU Wenjing ; FAN Yaofang ; TAN Xiaoying ; XU Yihuan ; WU Kun ; WEI Chuanchuan ; Key Laboratory of Freshwater Animal Breeding ; Ministry of Agriculture of P.R.C. ; Fishery College ; Huazhong Agricultural University
- 年：2016
- 作者机构：农业部淡水生物繁育重点实验室华中农业大学;
- 论文关键词：黄颡鱼 ; 脂代谢 ; PPARα ; 启动子 ; 非诺贝特
- 英文论文关键词：Pelteobagrus fulvidraco ; lipid metabolism ; PPARα ; promoter ; fenofibrate
- 会议召开时间：2016-11-02
- 会议录名称：2016年中国水产学会学术年会论文摘要集
- 语种：中文
- 分类号：S917.4
- 学会代码：OGSB
- 会议名称：2016年中国水产学会学术年会
- 会议地点：中国四川成都
- 主办单位：中国水产学会、四川省水产学会
- 学会名称：中国水产学会
- 页数：1
- 文件大小：588k
- 原文格式：D
- 会议级别：全国
摘要
PPARα是一类由配体激活的转录因子,属于核受体超家族成员,在调节糖脂代谢、细胞生长、分化和凋亡中发挥重要作用。近年来发现,对该转录因子在鱼类脂代谢调节中的研究较少。本实验以黄颡鱼为研究对象,通过克隆其PPARα基因的启动子序列,构建启动子缺失序列表达载体,并在非诺贝特的刺激下瞬时转染Hep G2细胞,确定PPARα启动子活性区域。再通过预测分析、突变等方法研究相关转录因子对PPARα表达的作用。结果显示,Deletion及重组转染后发现PPARα基因的启动子活性区域,在非诺贝特诱导下黄颡鱼PPARα启动子的转录活性显著增强;其次,在启动子区域上存在潜在的与脂代谢相关的转录因子结合位点,包括PPAR、HNF4、NF-kb、SP1等,将这些位点突变后转染发现启动子活性均降低。表明这些转录因子对调控黄颡鱼PPARα基因的表达有重要作用。本实验初步研究了在黄颡鱼脂代谢过程中如何对PPARα进行调控,为黄颡鱼脂类代谢调控提供了相关理论依据。
PPARα is a ligand-activated transcription factor belonging to the nuclear receptor superfamily, which plays a key role as regulators of glucose and lipid homeostasis as well as in cell proliferation, differentiation and apoptosis. Up to now, only limited information is available on regulation of lipid metabolism of PPARα promoter in fishes. In the present study, the promoter of PPARα was cloned from yellow catfish. Follwoing fenofibrate treatment, luciferase reporter gene recombinant plasmids were constructed and transfected into Hep G2 cell. Subsequently, we examined the promoter sequence, performed predictive and functional analyses to identify regulatory elements. Transient transfections detected the region that had higher promoter activity,the transcription activity of the promoter was observably increased after fenofibrate treatment.Furthermore, potential transcription factors binding sites related to lipid metabolism, such as PPAR, HNF4, NF-κb, SP1 etc., were identified. Additionally, promoter deletion analysis revealed that those factors are likely to have a crucial impact on the expression of PPARα.
PPARα is a ligand-activated transcription factor belonging to the nuclear receptor superfamily, which plays a key role as regulators of glucose and lipid homeostasis as well as in cell proliferation, differentiation and apoptosis. Up to now, only limited information is available on regulation of lipid metabolism of PPARα promoter in fishes. In the present study, the promoter of PPARα was cloned from yellow catfish. Follwoing fenofibrate treatment, luciferase reporter gene recombinant plasmids were constructed and transfected into Hep G2 cell. Subsequently, we examined the promoter sequence, performed predictive and functional analyses to identify regulatory elements. Transient transfections detected the region that had higher promoter activity,the transcription activity of the promoter was observably increased after fenofibrate treatment.Furthermore, potential transcription factors binding sites related to lipid metabolism, such as PPAR, HNF4, NF-κb, SP1 etc., were identified. Additionally, promoter deletion analysis revealed that those factors are likely to have a crucial impact on the expression of PPARα.
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