华西口腔医学杂志

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小鼠牙本质涎磷蛋白基因上游启动子的克隆和序列测定

郭 婷1,余 擎1,肖明振1,赵守亮1,高 杰1,朱庆林1,曹 罡2   

  1. 1.第四军医大学口腔医院 牙体科,陕西 西安710032;2.南京军区总医院 口腔科,江苏 南京210002
  • 收稿日期:2004-12-25 修回日期:2004-12-25 出版日期:2004-12-20 发布日期:2004-12-20
  • 通讯作者: 余 擎,Tel:029-83376073
  • 作者简介:郭 婷(1977-),女,河北人,住院医师,硕士
  • 基金资助:
    国家自然科学基金资助项目(30271418)

Cloning and Sequencing of the Upstream of Mouse Dentin Sialophosphoprotein Promoter

GUO Ting1,YUQing1,XIAO Ming-zhen1,ZHAO Shou-liang1,GAO Jie1,ZHUQing-lin1,CAO Gang2   

  1. 1.Dept.ofConservative Dentistry,Faculty ofSto- matology,The FourthMilitaryMedical University,Xi′an710032,China; 2.DentalDepartment ofNanjingMilitaryGeneralHos- pital,Nanjing210002,China
  • Received:2004-12-25 Revised:2004-12-25 Online:2004-12-20 Published:2004-12-20

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。

关键词: 牙本质涎磷蛋白, 聚合酶链式反应, 启动子

Abstract:

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.

Key words: dentin sialophosphoprotein, polymerase chain reaction, promoter