小鼠牙本质涎磷蛋白(DSPP)特异性启动子的克隆和序列分析

Cloning and sequence analysis of the specific promoter of mouse dentins ialophosphoprotein(DSPP)

目的 :获得小鼠牙本质涎磷蛋白 (DSPP)编码序列上游约 1.6kb的特异性启动子。方法 :按Mac Dougall报道设计一对引物 ,提取小鼠胚胎干细胞基因组DNA作为模板 ,PCR获得大小约为 1.6kb的DNA片段 ,将该片段克隆并进行序列分析。结果 :所得到的片段是DSPP的特异性启动子 ,其序列与文献报道有一定差异 ,但转录因子的结合位点均未发生突变。结论 :成功获得小鼠DSPP的特异性启动子 ,为进一步的研究打下基础。

Objective:To obtain the 1.6 kb distinct p romoter of mouse dentin s ialophosphoprotein(DSPP).Method: Primers for DSPP specific promo ter were design ed in accordance with the sequence reported by MacDougall et al, genomic DNA of mouse embryonic stem cell was extracted to be used as template in PCR, an approx imate 1.6 kb fragment was obtained, and it was then cloned and sequenced. Resul t: The fragment was confirmed to be DSPP specific promoter by partial sequenci ng, the sequencing result had some differences with that reported, but the bindi ng sites of transcription factors(such as Msx-1, AP1, SRE) were not found to be mutated.Conclusion: Mouse DSPP specific promoter was cloned succ essfully, which made basis for further studies. [