鸡肝脏型脂肪酸结合蛋白基因启动子活性分析  

Activity analysis of chicken liver fatty binding protein gene promoter

作　　者：高广亮[1,2] 冷丽[1,2] 张会丰[1,2] 贺綦[1,2] 李辉[1,2] 王启贵[1,3] 

机构地区：[1]东北农业大学农业部鸡遗传育种重点实验室,黑龙江哈尔滨150030 [2]东北农业大学动物科学技术学院,黑龙江哈尔滨150030 [3]重庆市畜牧科学院,重庆402460

出　　处：《中国兽医学报》2012年第9期1344-1348,共5页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(30972087)

摘　　要：PCR扩增鸡L-FABP基因5′侧翼区约2kb的DNA片段,进行克隆并测序,构建了鸡L-FABP基因报告基因系列缺失载体,瞬时转染进入人肝癌细胞系,利用双荧光素酶报告基因系统测定了荧光素酶活性。在线分析软件发现鸡L-FABP基因启动子区存在HNF-1、SREBP-1、AP-1、C/EBP、Oct-1、TATA、CCAAT、GATA-1等调控元件,没有发现CpG岛。报告基因结果表明鸡L-FABP基因启动子-2 076bp/-20bp区域具有最强的启动子活性,-522bp/-20bp区域启动子活性最弱;C/EBPα可以显著的抑制鸡L-FABP基因的表达,这些结果为深入研究鸡L-FABP的表达调控机制奠定了基础。The objective of this study was to analyze the promoter structure and activity of the chicken liver fatty binding protein（L-FABP） gene. The 5＇flanking 2 kb of chicken L-FABP gene was amplified by PCR,cloned and se- quenced. A series of recombination plasmids of L-FABP 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. Bioinformatics analysis revealed that the chicken L-FABP gene promoter fragment included HNF-1,SREBP- 1, AP-1, C/EBP, Oct- 1, TATA, CCAAT, GATA 1 and other regulatory elements binding sites and the gene promoter region can not find CpG islands. The activity of the promoter region analysis by luciferase reporter assays demonstra- ted that promoter region（--2 076 bp/--20 bp） was strongest promoter activity and the promoter region（--522 bp/ --20 bp） was worse,and C/EBPa could repress the chicken L-FABP gene expression. This study will provide the foundation for in-depth study on the chicken L-FABP gene expression regulation mechanism.

关 键 词：鸡 肝脏型脂肪酸结合蛋白基因 启动子 活性分析 

分 类 号：S852.2[农业科学—基础兽医学]