鸡肝脏胆汁酸结合蛋白基因启动子活性分析

Activity analysis of chicken liver bile acid binding protein gene promoter

研究对鸡肝脏胆汁酸结合蛋白基因(L-BABP)启动子活性进行研究。利用在线分析软件分析L-BABP基因启动子区,发现CCAAT box(-306)、TATA box(-995)、GATA-1(-1844)和SRE(-1866)等调控元件,没有发现CpG岛。扩增鸡L-BABP基因5'侧翼区约2 kb的DNA片段,构建鸡L-BABP基因报告基因系列缺失载体。报告基因结果表明,鸡L-BABP基因启动子-1 096/-66区域具有最强的启动子活性,-545/-62区域启动子活性最弱;C/EBPα可以显著抑制鸡L-BABP基因的表达,可为深入研究鸡L-BABP的表达调控机制奠定基础。

The objective of this study was to analyze the promoter structure and activity of the chicken liver bile acid binding protein （L-BABP） gene. Bioinformatics analysis revealed that the chicken L-BABP gene promoter fragment included HNF-1, SREBP-1, AP-1, C/EBP, Oct-l, TATA, CCAAT, GATA-1 and other regulatory elements binding sites and the gene promoter region cannot find CpG islands. The 5＇ flanking 2 kb of chicken L-BABP gene was amplified by PCR, cloned and sequenced. A series of recombination plasmids of L-BABP gene were constructed and transiently transfected into Human hepatoma cell line 2 （HEPG2）. Then their luciferase activity was measured by dual luciferase reporter gene assay system. The activity of the promoter region analysis by Luciferase reporter assays demonstrated that promoter region （-1 096/-66） was strongest promoter activity and the promoter region （-545/-62） was worse, and C/EBPa could repress the chicken L-BABP gene expression. This study will provide the foundation for in-depth study of the chicken L-BABP gene expression regulation mechanism.