转录因子Sp3与釉蛋白基因转录调控关系的研究

Relationship between Sp3 and the transcriptional regulation of enamelin gene

目的 探讨转录因子Sp3对釉蛋白基因（enamelin）的转录调控作用,以明确Sp3是否直接参与了釉蛋白基因的转录调控.方法 采用生物信息学软件预测釉蛋白基因上游调控区含有Sp3的结合位点;运用凝胶阻滞实验检测牙胚中的Sp3蛋白与釉蛋白调控区Sp3识别序列间的结合能力及其特异性;采用体外基因重组技术和双荧光素酶报告分析,将釉蛋白基因5＇调控区构建至pGL3-basic载体中,并将其与Sp3表达质粒体外共转染至永生化成釉细胞株（LS8）中,检测Sp3对釉蛋白基因的激活.结果 凝胶阻滞实验与双荧光素酶报告分析结果显示,Sp3不能直接结合在釉蛋白上游调控区并激活其转录.结论 Sp3没有以直接作用的方式参与釉蛋白基因的转录调控.

Objective To investigate the role of Sp3 in the transcriptional regulation of enamelin gene. Methods By bioinformatic analysis, a putative responsive element for Sp3 was identified.Electrophoretic mobility shift assay was used to examine the interaction between Sp3 and enamelin.5＇-flanking regulatory region of enamelin was cloned and ligated into pGL3-basic luciferase vector. Sp3 and the Enam-luc were cotransfected into mouse ameloblast-like cell line, and the activity of luciferase was examined. Results The results showed that Sp3 could not directly bind to the enamelin regulation region and activate enamelin transcription. Conclusions Sp3 might not be involved in transcriptional regulation of enamelin gene via an indirect interaction.