SPDEF调控小鼠成釉细胞MMP-20基因表达的分子机制研究  

Study of Transcription Factor SPDEF on Regulating the Expression of Matrix Metalloproteinase-20

作　　者：纪虹利[1] 杜森[1] 董超[1] 张世龙[1] 靖旭[1] 高玉光[2] 

机构地区：[1]潍坊医学院口腔医学研究所,山东潍坊261053 [2]滨州医学院口腔医学院

出　　处：《潍坊医学院学报》2013年第1期18-21,共4页Acta Academiae Medicinae Weifang

基　　金：国家自然科学基金资助课题(基金编号:30973327);山东省自然科学基金资助课题(基金编号:(ZR2010HM076)

摘　　要：目的通过克隆转录因子SPDEF以及其真核表达载体的构建,分析该基因对成釉细胞MMP-20基因表达的影响,为进一步研究转录因子SPDEF在牙釉质形成中发挥的作用奠定基础。方法利用RT-PCR技术从小鼠成釉细胞中获得SPDEF基因,测序正确后克隆至pcDNA3.1/myc-HisA载体中,将重组质粒瞬时转染成釉细胞,并用双荧光素酶报告基因系统检测技术来检测MMP-20启动子区域的转录活性。结果①SPDEF基因经过RT-PCR扩增得到的产物与预期片段978bp相符,将所获得的目的基因与GenBank提供的已知序列(NM:013891.4)完全一致。②重组质粒pcDNA3.1/myc-HisA-SPDEF转染成釉细胞后,用荧光素酶分析系统检测显示MMP-20启动子转录活性升高。结论成功构建了SPDEF真核表达载体,初步研究证明SPDEF可能与MMP-20特征性序列结合从而上调MMP-20的表达。Objective To analyse the influence of the expression of MMP-20 gene by cloning the transcription faetor SPDEF gene and constructing eukaryotie expression vector in order to further study the role of transcription factor SPDEF in the fi）rmation of the enamel. Methods The SPDEF gene was acquired from ALC by RT-PCR,Then inserted into pcDNA3. 1/myc-HisA vector. The recombinant plasmid was transiently Iransfeeted into ALC. Dual-Luciferase Reporter Assay System was pefl＇onned to deteet the traqseriptional activity of MMP-20 lU＇umoter region. Results ①SPDEF was amplification by RT-PCR and matehed the expeeted fragment about 978hp. The sequencing result was completely matched wilh the known sequence in GeneBank accession NO. NM ：013891.4. ②The recombinant plasmid of pcDNA3. l/mye-SI＇l）EF was transfect into ALC. Then detect the increase of the transcriptional activity of MMP-20. Conclusion The eukaryotic ex pression vector carried SPI）EF is constructed successfully. Preliminary analysis has shown that SPDEF may be combined with the eharacteristic sepuence of MMP-20 which up-regulate the expression of MMP-20.

关 键 词：转录因if-SPDEF 金属机制蛋白 -20 RT—PCR 双荧光素酶报告基因检测 

分 类 号：R392.13[医药卫生—免疫学]