小鼠牙本质涎磷蛋白基因上游启动子的克隆和序列测定

目的　克隆并测定小鼠牙本质涎磷蛋白(DSPP)基因上游启动子的序列。方法　提取成年Balb/c鼠基因组DNA,设计引物,通过聚合酶链反应(PCR)得到小鼠牙本质涎磷蛋白基因上游启动子的序列。再将目的片段定向连入T载体,酶切鉴定并测序。结果　将小鼠牙本质涎磷蛋白基因上游启动子序列分3段克隆,分别得到997 bp、1 004 bp及674 bp大小的目的片段。连入载体后,酶切结果测定重组质粒成功。其测序结果与小鼠基因组染色体位置5q21处的相应序列99%一致。结论　成功克隆到牙本质涎磷蛋白基因上游启动子的序列,为进一步研究DSPP基因的转录调控机制奠定了基础。

Cloning and Sequencing of the Upstream of Mouse Dentin Sialophosphoprotein Promoter

Objective To clone and sequence the upstream of mouse dentin sialophosphoprotein promoter.Methods Genomic DNA was obtained from Balb/c mouse blood. The upstream of mouse dentin sialophosphoprotein promoter segments was obtained by PCR. Then the segments were inserted into T-vector. The plasmids were identified by digestion with the restriction enzyme analysis. The positive clone was sequenced and compared with Genebank.Results The upstream of mouse dentin sialophosphoprotein promoter was divided into three sequences and three different target segments with 997 bp, 1 004 bp and 674 bp in length were obtained. After identified, sequenced and compared with Genebank, the sequences of the segments were consistent with those displayed on Genebank by 99%.Conclusion The clone of the upstream of mouse dentin sialophosphoprotein promoter was successful. This work will help to study the regulation of DSPP expression.