【摘要】 目的:研究小鼠Lrp5基因基本启动子。方法:PCR扩增小鼠Lrp5基因基本启动子序列,构建荧光素酶报告基因表达体系,以PRL鄄TK为内参照质粒,瞬时转染COS鄄7细胞熏48h后收集细胞测定荧光素酶相对表达活性。结果:在小鼠Lrp5基因基本启动子区域,构建了三种荧光素酶报告基因表达体系pGL3鄄16(鄄16bp～+132bp)、pGL3鄄54(鄄54bp～+132bp)和pGL3鄄103(鄄103bp～+132bp)。pGL3鄄103表达载体的相对荧光素酶活性最高;pGL3鄄54的相对荧光素酶活性是pGL3鄄103表达载体的80%;而pGL3鄄16相对荧光素酶表达活性急剧下降,与阴性对照pGL3鄄basic的活性相近,无启动子活性。结论:鄄54bp～鄄16bp区域内含有小鼠Lrp5基因转录所必需的基本启动子序列,其中三个SP1保守序列是小鼠Lrp5基因基本启动子所必需的。更多还原

【Abstract】 Objective: To study the basal promoter of mouse Lrp5 gene. Methods: The region supposed to contain the basal promoter of mouse Lrp5 gene was amplified by PCR, and luciferase expression vectors were constructed. These expression vectors were transfected into COS-7 cell line and the relative luciferase activity was measured 48 h later. Results: Three luciferase expression vectors, pGL3-16(-16 bp～+132 bp), pGL3-54(-54 bp～+132 bp) and pGL3-103(-103 bp～+132 bp) were constructed. The relative luciferase activity of pGL3-103 was the highest, the activity of pGL3-54 was 80% of the highest activity, and the activity of pGL3-16 was significantly lower than those of pGL3-103 and pGL3-54 deeply. Conclusion: The region from-54 to-16 bp includes an essential promoter sequence for mouse Lrp5 gene transcription. The region contains three SP1 elements, which are necessary for the transcription of mouse Lrp5 gene.更多还原