作　　者： ; ; ; ; ; ;

机构地区： 第三军医大学基础部微生物学教研室重庆市微生物工程重点实验室

出　　处： 《第三军医大学学报》 2006年第9期896-899,共4页

摘　　要： 目的对IRF-3的主要靶基因———小鼠IFN-β基因启动子区域进行分析并构建IRF-3报告基因载体。方法将小鼠IFN-β基因启动子全序列以及不同截短片段克隆到无启动子的pGF3basic/enhancer质粒中,以EGFP为报告基因,转染小鼠Raw264.7巨噬细胞系后观察细胞对LPS刺激的反应。结果IFN-β启动子PRD调控区域上游序列、PRD I区,PRD II区和PRD IV区缺失后,LPS同样能诱导EGFP报告基因的表达。只保留PRD区及IFN-β启动子核心区的载体转染Raw264.7细胞后,可以在LPS刺激下诱导EGFP报告基因的高表达。结论构建了能检测IRF-3活性的报告基因载体,为进一步研究LPS激活IRF-3的信号转导通路奠定了基础。 Objective To analyze the regulatory elements that regulates the transcription of murine IFN-β gene and to construct IRF-3 reporter gene. Methods The promoter region of murine IFN-β gene was from murine genomic DNA by PCR and was cloned into promoter-free plasmid pGF3basic/enhancer, amplified upstream of EGFP reporter gene. Vectors with serial deletions at 5＇ side of promoter region were also constructed. Following transient transfection of murine Raw264.7 cells with the vectors and LPS stimulation, the expression of EGFP reporter gene was observed under fluorescent microscopy. Results This study showed that LPS could induce expression of EGFP reporter gene when DNA fragment upstream of PRD region, as well as PRD Ⅰ , Ⅱ and Ⅳ region, were deleted. Conclusion An IRF-3 EGFP reporter gene was established by including only PRD Ⅲ sequence and core region of IFN-β promoter. This reporter gene is an important tool for investigation of LPS-induced IRF-3 signaling transduction pathways.

关 键 词： 启动子 调控元件 报告基因

领　　域： [生物学] [生物学]