小鼠牙本质涎磷蛋白基因上游启动子的克隆和序列测定
Cloning and Sequencing of the Upstream of Mouse Dentin Sialophosphoprotein Promoter
作者：郭婷,余擎,肖明振,赵守亮,高杰,朱庆林,曹罡
Author：GUO Ting1,YUQing1,XIAO Ming-zhen1,ZHAO Shou-liang1,GAO Jie1,ZHUQing-lin1,CAO Gang2
收稿日期：2004-12-25 年卷（期）页码：2004,22(06):513-
期刊名称：华西口腔医学杂志
Journal Name：West China Journal of Stomatology
关键字：牙本质涎磷蛋白,聚合酶链式反应,启动子,
Key words：dentin sialophosphoprotein,polymerase chain reaction,promoter,
基金项目：国家自然科学基金资助项目(30271418)
中文摘要
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。
英文摘要
Objective To clone and sequence the upstream of mouse dentin sialophosphoprotein promoter.Methods Genom- ic DNAwas obtained from Balb/c mouse blood. The upstream of mouse dentin sialophosphoprotein promoter segmentswas 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 sialophospho- protein 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 comparedwithGenebank, the sequences of the segmentswere consistentwiththose displayed on Genebank by 99%.Conclusion The clone of the upstream of mouse dentin sialophosphoprotein promoterwas suc- cessful. This work will help to study the regulation of DSPP expression.
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