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牛脂肪酸结合蛋白4基因5′调控区的组织表达分析、克隆及特征研究

Tissue expression analysis, cloning, and characterization of the 5'-regulatory region of the bovine fatty acid binding protein 4 gene.

作者信息

Li A, Zhao Z, Zhang Y, Fu C, Wang M, Zan L

出版信息

J Anim Sci. 2015 Nov;93(11):5144-52. doi: 10.2527/jas.2015-9378.

Abstract

Fatty acid binding protein 4 (FABP4) is a member of the FABP family of proteins that bind fatty acids and play important roles in fatty acid uptake and intracellular transport. In the present study, we cloned the 5'-regulatory region of bovine and identified its transcription initiation sites. Sequence comparative analysis demonstrated amino acids and promoter sequences of were highly conservative in mammals. Real-time PCR analysis revealed the products of bovine were very highly expressed in subcutaneous adipose tissue. Serial deletion constructs of the bovine 5'-regulatory region evaluated in a dual-luciferase reporter assay showed that 209 bp upstream from the transcription initiation site was its core promoter. An electrophoretic mobility shift assay combined with a site-directed mutation experiment indicated that peroxisome proliferator activated receptor gamma (PPARγ), CCAAT/enhancer-binding protein β (C/EBPβ), and sterol regulatory element-binding protein (SREBP) were important transcription factors for bovine . These results provide an important basis for further understanding the regulation of bovine .

摘要

脂肪酸结合蛋白4(FABP4)是脂肪酸结合蛋白家族的成员,该家族蛋白可结合脂肪酸并在脂肪酸摄取和细胞内运输中发挥重要作用。在本研究中,我们克隆了牛的5'-调控区并确定了其转录起始位点。序列比较分析表明,其氨基酸和启动子序列在哺乳动物中高度保守。实时定量PCR分析显示,牛的产物在皮下脂肪组织中高表达。在双荧光素酶报告基因分析中评估的牛5'-调控区的系列缺失构建体表明,转录起始位点上游209 bp是其核心启动子。电泳迁移率变动分析结合定点突变实验表明,过氧化物酶体增殖物激活受体γ(PPARγ)、CCAAT/增强子结合蛋白β(C/EBPβ)和固醇调节元件结合蛋白(SREBP)是牛的重要转录因子。这些结果为进一步了解牛的调控提供了重要依据。

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