猪THRSP基因5’调控区多位点SNP联合效率的研究

Study on the promoter efficiency of multi-sites SNP in 5' regulatory region of THRSP gene in pigs

旨在分析猪THRSP基因5’调控区多位点SNP联合调控THRSP基因表达的效率。实验从猪基因组获取包含多位点SNP的目的片段,利用荧光素酶报告基因pGL3-Basic构建表达载体,检测多位点SNP的联合启动效率,并通过实时荧光定量PCR进行THRSP基因表达验证分析。结果表明,在5个SNP位点中,TCAGA的启动效率最高,比CTGGA的启动效率高129%,而CCAGA、CTAGA和CTGAT的启动效率较CTGGA分别低99.0%、73.2%和42.6%。TCAGA的启动效率较CCAGA高75.6%,而CCAGA比CTAGA的启动效率高96.6%;报告基因所揭示的SNP启动效率与THRSP基因实时荧光定量PCR结果一致。

This paper aims to analyze the promoter efficiency of multi-sites SNP to THRSP gene expression in the 5＇ regulatory region of pig THRSP gene. The fragments containing 5 SNP sites were obtained from the 5＇ regulatory region in pig genome. The pGL3-Basic expression vectors of luciferase reporter gene were constructed with the multi-sites SNE THRSP gene expression was detected by real-time quantitative PCR. The results showed that the TCAGA promoter efficiency in five SNP loci was highest, which increased 129% than that of CTGGA. Compared with CTGGA, the promoter efficiency of CCAGA, CTAGA and CTGAT were lower 99.0%, 73.2 % and 42.6 %, respectively. The promoter efficiency of TCAGA increased 75.6% than that of CCAGA, while CCAGA increased 96.6% than that of CTAGA. The promoter efficiency of SNP was revealed by reporter gene as the results by real-time PCR gene for THRSP gene expression