摘要： 旨在初步分析猪EIF2S3基因的启动子活性以及转录调控元件。利用生物信息学分析、PCR扩增、基因克隆、细胞转染、双荧光素酶活性分析等方法，获得了EIF2S3基因启动子区域的序列特征，构建了不同片段长度的EIF2S3基因启动子区的双荧光素酶报告基因载体并分析了其荧光素酶活性，进而确定了EIF2S3基因的核心启动子区域以及关键的调控区域，最后还预测了关键调控区域的转录因子及其结合位点。结果表明，EIF2S3基因候选启动子区包含3个核心启动子以及2个CpG岛；-706~+200 bp为EIF2S3基因的核心启动子区域，-706~-253 bp为EIF2S3基因启动子的关键调控区域，且发挥正向调节作用，而-1 563~-706 bp不存在任何对EIF2S3基因启动子活性有影响的调控元件；EIF2S3基因启动子的关键调控区域包含440个转录因子结合位点，且多个转录因子在此区域均有多个结合位点，如SP1、KLF4、MyoD1、MyoG、NFKB1。为进一步研究猪EIF2S3基因的表达机制奠定了基础。

关键词: 猪, EIF2S3, 启动子, 转录活性, 转录因子

Abstract: The purpose of this study was to initially analyze the promoter activity and transcriptional regulatory elements of porcine EIF2S3 gene.Using the bioinformatics analysis,PCR amplification,gene clone,cell transfection and dual luciferase activity assay.We obtained the sequence characteristics of the promoter region of EIF2S3,constructed dual luciferase reporter gene vector with different fragment lengths in the EIF2S3 promoter region and analyzed its luciferase activity,identified the core promoter region and key regulatory regions of the EIF2S3,and predicted transcription factors and their binding sites in key regulatory regions.The results showed that the candidate promoter region of EIF2S3 contained 3 core promoters and 2 CpG islands.The -706~+200 bp was the core promoter region and the -706~-253 bp was the key regulatory region of EIF2S3 promoter.This key regulatory region played a positive regulatory role in the transcriptional regulation of EIF2S3 gene.There was no regulatory element that affects the promoter activity in the -1 563~-706 bp of EIF2S3 promoter.The key regulatory region of EIF2S3 promoter contained 440 transcription factor binding sites,and many transcription factors in this region had multiple binding sites,such as SP1,KLF4,MyoD1,MyoG,NFKB1.These data provided a basis for studying the expression mechanism of pig EIF2S3.

Key words: Pig, EIF2S3, Promoter, Transcriptional activity, Transcription factor

中图分类号: 

• S828.2

• Q78