Zhao Zhidong, Tian Hongshan, Shi Bingang, Jiang Yanyan, Liu Xiu, Hu Jiang
College of Animal Science and Technology & Gansu Key Laboratory of Herbivorous Animal Biotechnology, Gansu Agricultural University, Lanzhou 730070, China.
Animals (Basel). 2019 Sep 5;9(9):654. doi: 10.3390/ani9090654.
Oleic acid is a major monounsaturated fatty acid, which accounts for about 33% of the fatty acid content in beef and is considered to have the least negative effect on serum cholesterol levels. Fatty acid transport protein 1 (FATP1), an integral membrane protein that facilitates long-chain fatty acid (LCFA) influx, is involved in the genetic network for oleic acid synthesis in beef. Its expression exhibits significant positive correlations with intramuscular fat (IMF) content in the longissimus thoracis. However, the expression mechanism of or is still unclear. To elucidate the molecular mechanisms involved in bovine regulation, we cloned and characterized the promoter region of . By applying 5'-rapid amplification of cDNA end analysis, we identified two alternative splice variants of this gene. Using a series of 5' deletion promoter plasmids in luciferase reporter assays, we found that the core promoter was 96 base pairs upstream from the transcription initiation site. Electrophoretic mobility shift assay combined with a site-directed mutation experiment demonstrated that KLF15 binding to the promoter region drives the transcription. KLF15 plays an essential role in adipogenesis and skeletal muscle lipid flux. Thus, these results might provide further information on the regulatory roles of gene in mediating the lipid composition in beef.
油酸是一种主要的单不饱和脂肪酸，约占牛肉脂肪酸含量的33%，被认为对血清胆固醇水平的负面影响最小。脂肪酸转运蛋白1（FATP1）是一种促进长链脂肪酸（LCFA）内流的整合膜蛋白，参与牛肉油酸合成的遗传网络。其表达与胸最长肌的肌内脂肪（IMF）含量呈显著正相关。然而，FATP1的表达机制仍不清楚。为了阐明牛FATP1调控的分子机制，我们克隆并鉴定了FATP1的启动子区域。通过应用5'-cDNA末端快速扩增分析，我们鉴定出该基因的两个可变剪接变体。在荧光素酶报告基因检测中使用一系列5'缺失启动子质粒，我们发现核心启动子位于转录起始位点上游96个碱基对处。电泳迁移率变动分析结合定点突变实验表明，KLF15与启动子区域的结合驱动了FATP1的转录。KLF15在脂肪生成和骨骼肌脂质通量中起重要作用。因此，这些结果可能为FATP1基因在调节牛肉脂质组成中的作用提供进一步的信息。
