畜牧兽医学报 ›› 2010, Vol. 41 ›› Issue (6): 651-656.doi:

• 遗传繁育 • 上一篇    下一篇

鸡脂蛋白酯酶基因启动子的克隆及活性分析

毕静,丁宁,王宁,李辉*   

  1. 东北农业大学动物科学技术学院,哈尔滨 150030
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2010-06-25 发布日期:2010-06-25
  • 通讯作者: 李辉

Cloning and Activity Analysis of Promoter of Chicken Lipoprotein Lipase Gene

BI Jing, DING Ning, WANG Ning, LI Hui*   

  1. College of Animal Science and Technology, Northeast Agricultural University, Harbin 150030, China
  • Received:1900-01-01 Revised:1900-01-01 Online:2010-06-25 Published:2010-06-25

摘要: 本文旨在研究鸡脂蛋白酯酶基因(lipoprotein lipase, LPL)启动子的结构和启动子活性。采用PCR方法扩增了鸡LPL基因5′侧翼区2 kb的DNA片段,对其进行克隆、测序及序列分析后,构建了其全长及系列截断突变的报告基因表达载体,瞬时转染鸡胚成纤维细胞(DF1),用双荧光素酶报告系统测定了荧光素酶活性。生物信息学分析发现,鸡LPL基因启动子区存在Oct-1、GC box、CCAAT、GATA、AP1等调控元件,在启动子-575~+137 bp区域内存在一个CpG岛。报告基因分析表明,鸡LPL基因的启动子-359~+163 bp区域就具有启动子活性,启动子-601~+163 bp区域具有最强的启动子活性。结果显示,鸡LPL基因受多种转录因子和上游序列的调控,本研究为深入研究鸡LPL基因的表达调控机制奠定了基础。

Abstract: The objective of this study was to analyse the promoter structure and activity of the chicken lipoprotein lipase (LPL) gene. 2 kb 5′ flanking region of chicken LPL gene was amplified by PCR, cloned and sequenced. Subsequently, the luciferase reporter gene plasmids containing LPL gene promoter and its serial promoter deletions were constructed and introduced into chicken fibroblast cell line DF-1. The expression of luciferase was measured in transient expression assays. Bioinformatics analysis showed that the promoter fragment contained binding sites of transcription factors Oct1, GC box, CCAAT, GATA, AP1 and it also had a CpG island from -575 to +137 bp. Luciferase reporter assays demonstrated that the promoter region from -359 to +163 bp confered basal transcriptional activity and the region from -601 to +163 bp confered the most powerful transcriptional activity of the chicken LPL gene. These results indicated that chicken lipoprotein lipase gene could be regulated by variety of transcription factors and upstream elements. This study pays the way for further research on the regulation mechanism of chicken LPL gene.