摘要： 旨在克隆和分析猪STAB2基因启动子及其转录活性。利用基因克隆、双荧光素酶报告基因系统以及生物信息学分析等方法，克隆得到了STAB2基因转录起始位点上游1 997 bp的候选启动子序列并对其序列特征进行了分析，同时，构建了不同长度的5'端缺失的重组载体并利用双荧光素酶报告基因系统分析了其荧光素酶活性，进而确定了STAB2基因的核心启动子区域及关键调控区域，并对关键调控区域内的转录因子及其结合位点进行了分析。结果表明：STAB2基因的候选启动子区域内包含4个核心启动子和1个CpG岛；-309--39 bp为STAB2基因的关键调控区域，-1 045--309 bp可能存在一个正向调控元件，而-1 506--1 045 bp可能存在一个负向调控元件；STAB2基因的关键调控区域内包含72个转录因子结合位点，部分转录因子在这一区域具有多个结合位点，如Arnt：：Ahr、ZNF354C、Klf4和KLF5，并且，转录因子结合位点之间也存在重合区。为进一步研究猪STAB2基因的表达调控机制以及其在调控猪抗病和肌肉品质中的功能提供了理论依据。

关键词: 猪, STAB2, 启动子, 克隆, 转录活性

Abstract: The aim of these study was to clone the promoter of porcine STAB2 gene and analysis its transcriptional activity. Using the gene cloning, dual luciferase reporter gene system and bioinformatics analysis methods, we cloned a 1 997 bp candidate promoter sequence of STAB2 gene and analysed its sequence characteristics, constructed various recombinant vectors with 5' deletions and analysed its luciferase activity by the dual luciferase reporter gene system. Thus, we identified the core promoter region and key regulatory regions of STAB2 gene. We also analysed the transcription factors and their binding sites in the key regulatory regions of STAB2 gene. The results showed that the candidate promoter regions of STAB2 gene contained four core promoters and one CpG island. The-309--39 bp was the key regulatory regions of STAB2 gene's promoter. There was a positive regulatory element in the-1 045--309 bp and a negative regulatory element in the-1 506--1 045 bp of STAB2 gene's promoter. The key regulatory regions of STAB2 gene contained 72 transcription factor binding sites, and some transcription factors had multiple binding sites in this regions, such as Arnt：：Ahr, ZNF354C, Klf4 and KLF5. We also found that coincident region existed between transcription factor binding sites. This study can provide a theoretical basis for further study of the expression regulation mechanism of STAB2 gene and its function in regulating disease resistance and muscle quality in pigs.

Key words: Pig, STAB2, Promoter, Clone, Transcriptional activity

中图分类号: 

• S828.2