速激肽受体1基因启动子近端E盒点突变对下游基因调控的影响  

Effect of point mutation in the upstream proximate E-box of neurokinin receptor 1 gene on downstream gene expression

作　　者：陈玲群 王练红[1] 周金[1] 王林 

机构地区：[1]怀化市第二人民医院检验科,怀化418000 [2]湖南医药学院检验系

出　　处：《山西医科大学学报》2015年第6期550-552,共3页Journal of Shanxi Medical University

摘　　要：目的研究NK1R转录起始位点上游启动子近端E盒点突变对下游基因转录活性的影响。方法定位NK1R基因转录起始点上游近端E盒结构,克隆含近端E盒的启动子序列,对E盒进行点突变,构建野生型和突变型荧光素酶报告基因载体,分别与内参荧光素酶报告基因载体p GL4.74 hluc转染MCF-7细胞,根据转染质粒类型分为PGL3野生型组、PGL突变型组、PGL3-basic组。结果 PGL3野生型组NK1R荧光素酶载体相对表达量为0.75±0.05;PGL3突变型组荧光素酶载体NK1R相对表达量0.60±0.07;PGL3-basic组相对表达量为0.38±0.05。结论 E盒结构可以影响所在启动子对下游基因的转录激活作用,点突变可以降低对下游基因的转录活性。Objective To investigate the influence of point mutation in upstream proximate E-box of neurokinin receptor 1 gene on down- stream gene expression. Methods The E-box proximate upstream end was located, and the promoter sequence containing E-box was cloned. E-box was mutated by overlapping PCR, and the wild promoter and mutation promoter luciferase reporter vectors were construc- ted. The constructed vectors were transfected into MCF-7 cells. The experiment included three groups： PGI3 wild group,PGL3 mutation group, and PGL3 basic group. Results The related luciferase expression was 0.75 ± 0.05 in PGL3 wild group, 0.60 ± 0.07 in PGL3 mutation and 0.38 ± 0.05 in PGL3 basic group. Conclusion E-box can regulate the downstream gene expression, and the point muta- tion in E-box downregulate the transcription activity of downstream genes.

关 键 词：速激肽受体1 E盒 启动子 荧光素酶报告基因载体 

分 类 号：Q754[生物学—分子生物学]