中国医科大学学报2019，Vol.48Issue(1)：29-33,5.DOI:10.12007/j.issn.0258-4646.2019.01.006
小鼠Na+-K+-2Cl-共转运蛋白基因启动子克隆及20-HETE对其转录活性的影响
Cloning of the Murine Na+-K+-2Cl-Cotransporter Gene Promoter and the Effect of 20-HETE on Its Transcriptional Activity
摘要
Abstract
Objective To clone the murine Na+-K+-2Cl-cotransporter (Nkcc2) gene promoter and analyze 20-HETE regulation of the murine Nkcc2 gene transcriptional activity. Methods A fragment of the murine Nkcc2 gene promoter was analyzed using bioinformatics software AliBaba and TRANSFAC TESS. The murine Nkcc2 gene promoter fragment (-1 462 bp-+40 bp) was amplified by PCR using murine genomic DNA as a template and then cloned into a pGL3-Basic vector to generate a luciferase reporter construct. The recombinant reporter construct was transiently transfected into HEK293 T cells using Lipofectamine 2000 for 24 h. The transfected HEK293 T cells were treated with 20-HETE for 2 h followed by measurement of the luciferase activity using the Dual-Luciferase Reporter Assay system. Results A luciferase reporter construct containing the murine Nkcc2 gene promoter was successfully generated. The results showed that 20-HETE significantly reduced the transcriptional activity of the construct. Conclusion 20-HETE may reduce the expression of the murine Nkcc2 gene through transcriptional regulation.关键词
Na+-K+-2Cl-共转运蛋白/启动子/20-HETE/基因克隆Key words
Na+-K+-2Cl-cotransporter/promoter/20-HETE/gene clone分类
医药卫生引用本文复制引用
武晶晶,孔令慧,贾茹..小鼠Na+-K+-2Cl-共转运蛋白基因启动子克隆及20-HETE对其转录活性的影响[J].中国医科大学学报,2019,48(1):29-33,5.基金项目
国家自然科学基金青年科学基金 (81400709) （81400709）
