中文摘要：

目的 ：克隆小鼠干扰素基因刺激因子（STING）基因启动子,并对其启动子活性和转录调控机制进行初步探讨。方法 ：利用PCR方法扩增小鼠STING基因5′上游1 005 bp（-927~＋77）的片段,亚克隆至p GL3-basic质粒,然后通过步移缺失构建不同缺失片段的重组质粒。双荧光素酶报告活性分析检测重组质粒在NIH3T3中的活性,并利用生物信息学方法预测转录因子结合位点。结果：经酶切、测序鉴定,成功构建了小鼠STING启动子荧光素酶报告基因重组质粒。与p GL3-basic质粒相比,STING启动子重组质粒的相对荧光素酶活性增加（P〈0.05）。通过生物信息学软件预测小鼠STING启动子区域（-177~-48）可能含有GATA、IK2、MZF1、SP1/SP3、STAT等转录因子结合位点。结论：成功构建小鼠STING不同缺失片段的启动子荧光素酶报告基因重组质粒。通过活性比较,推测小鼠STING的核心启动子区位于-177~＋77区域,其中可能含有多个潜在的转录因子结合序列。

英文摘要：

Objective：To clone the promoter sequences of stimulator of interferon gene（STING） and evaluate its activity, and to preliminarily investigate the transcriptional regulatory mechanisms. Methods： Promoter region was predicted by bioinformatics methods, and the 1 005 bp（-927-＋77） fragment of 5＇ upstream sequences of STING gene was amplified by PCR, and then cloned to pGL3-basic vector to construct the luciferase report gene recombinant plasmid. Three promoter fragments with different length were obtained by walking deletion and cloned into pGL3-basic vector. The vector expression activities were determined by transfection of the mouse NIH3T3 cells with the recombinant plasmids of STING gene promoter. Bioinformatics methods were performed to predict the potential transcriptional factor binding sequences. Resluts： The luciferase reporter gene recombinant vectors of mouse STING promoter were successfully constructed. Compared with the pGL3-basic plasmid, the relative luciferase activities of recombinant vectors of STING promoter were much higher （P 〈 0.05）. In addition, the binding sequences of GATA, IK2, MZF1, SP1/SP3, and STAT may be included in the promoter region （-177~-48） of STING gene,which were predicted by the bioinformatics method. Conclusion： The luciferase report gene recombinant plasmids of STING gene promotor were constructed successfully and had strong transcriptional activity in NIH3T3 cells. By the activity comparison, it is speculated that the core promoter region of mouse STING is located in the -177--＋77 region, which may contain a number of potential transcription factor binding sequences.