人牙釉质蛋白Amelotin,Ameloblastin及Enamelin基因启动子的初步研究  

Preliminary Study of Human Enamel Protein Amelotin,Ameloblastin and Enamelin Gene Promoter

作　　者：刘晓影[1] 高志芹[1] 韩婷婷[2] 官秀梅[3] 高玉光[2] 

机构地区：[1]潍坊医学院细胞生物学教研室,山东潍坊261053 [2]潍坊医学院口腔研究所 [3]潍坊医学院生化教研室

出　　处：《潍坊医学院学报》2009年第5期321-326,共6页Acta Academiae Medicinae Weifang

基　　金：国家自然科学基金资助项目(课题编号:30672316);山东省自然科学基金资助项目(课题编号:Y2006C106)

摘　　要：目的分析人釉成熟蛋白(Amelotin,AMTN)、成釉蛋白(Ameloblastin,AMBN)与釉蛋白(Enamelin,ENAM)基因上游启动子序列,构建不同长度的基因上游启动子报告基因载体,为进一步判断启动子序列的转录调控区奠定基础。方法利用软件分析基因启动子序列,以PCR方法获取基因上游启动子片段,将其构建至报告基因载体pGL3-Basic中。结果成功地获得了不同长度的Amelotin,Ameloblastin及Enamelin基因启动子目的片段,酶切鉴定表明不同长度启动子荧光素酶报告基因载体构建成功。结论成功构建了不同长度启动子的pGL3-Basic荧光素酶报告基因载体,为进一步研究牙釉质蛋白基因启动子的转录活性及转录调控特点奠定基础。Objective Analyzing upstream promoter sequence of human Amelotin （ AMTN ） , human ameloblastin（AMBN） and human ameloblastin（ENAM） gene, constructing luciferase report gene vectors with different promoter segments and so as to provide a foundation for confirming transcription regulation district of promoter sequence in further study. Methods Software was used to analysis gene promoter sequence. Different-length upstream promoter segments of Amelotin, Ameloblastin and Enamelin gene were obtained through PCR method using recombinant T vector as template, and than these segments were cloned into luciferase report gene vectors pGL3-Basic. Results Different-length promoter segments of Amelotin, Ameloblastin and Enamelin gene were obtained. The different-length promoter recombinant luciferase reporter vectors were constructed successfully appraised by cutting them with two different restrict enzymes. Conclusion The successful construction of different-length recombinant pGL3-Basic luciferase reporter gene vector,which will provide a foundation for further studying the transcriptional activity and characteristics of transcriptional regulation of enamel protein gene promoter.

关 键 词：Amelotin AMELOBLASTIN ENAMELIN pGL3-Basic荧光素酶报告基因载体 

分 类 号：R318[医药卫生—生物医学工程]