人EPHB2基因启动子的初步鉴定

Characterization of Promoter of Human EPHB2 Gene

目的为进一步研究人EPHB2基因的表达调控机制,初步鉴定、分析该基因的启动子。方法在对人EPHB2基因的生物信息学分析的基础上,在EPHB2基因的转录起始位点已明确的前提下,克隆了该基因的5′侧翼区,并对该区进行功能分析,分别构建了5种含不同长度启动子序列的荧光素酶报告基因表达质粒,将它们瞬时转染293细胞,检测其荧光素酶活性。结果EPHB2的5′侧翼区的-138～+83区域的启动子活性不高,而从-425～+83开始显著升高,-1174～+83区域的启动子活性又出现明显降低。结论人EPHB2基因启动子的较小区域位于其5′侧翼区的-425～-139区域。另外,在-1174～-970区域可能存在着沉默子。

Objective To identify and analyze the promoter of EPHB2 gene. Methods The 5＇-flanking region of EPHB2 gene was cloned. The region was analyzed by using bioinformatic approach, and five luciferase expression vectors containing deleted human EPHB2 gene promoter were constructed. All expression vectors were transfected into 293 cells. Results The relative promoter activity of the -138 to ＋ 38 region was not high, but it began to increase from the -425 to ＋ 83 region. From the region of-1 174 to ＋ 83, it was significantly decreased. Conclusion The luciferase assays revealed that the -425 to -139 region was the promoter of the human EPHB2 gene. Moreover, there may be silent elements in the -1 174 to -970 region.