摘要
目的:研究Cbfαl对釉原蛋白启动子转录活性的影响。方法:利用生物信息学的方法分析釉原蛋白启动子区域的序列,确定可能存在的结合位点,将利用PCR及酶切的方法从小鼠C57BL/6J基因组上扩增的、不同长度的釉原蛋白启动子构建的虫荧光素酶报告载体与Cbfαl共转Hela细胞,分析Cbfαl对不同长度的启动子转录活性的影响,将包含整个启动子区域的报告载体与Cbfαl共转Hela、CHO、UMR-106细胞,分析在不同细胞中Cbfαl对全长启动子转录活性的影响。结果:在Hela细胞中,Cbfαl对各段启动子都有激活作用,且该作用不随着启动子长度的变化而变化,在CHO、UMR-106细胞中,Cbfαl对启动子转录活性的影响很弱。结论:在Hela细胞中,Cbfαl对釉原蛋白启动子的转录活性有明显的增强作用,而且在对逐步缩短的启动子表现出一致的增强效能。结合生物信息学对Cbfαl结合位点的分析,可以初步判断在启动子下游,第一个内含子中存在Cbfαl的作用位点。另外,这种增强作用表现出明显的上皮细胞特异性,表明Cbfαl对釉原蛋白启动子的增强作用是有组织特异性的。
AIM:To investigate the role of Cbfαl in the regulation of Amelogenin expression.METHODS:Firstly,the 5′ flanking of Amelogenin gene was anylized,and some potential Cbfαl binding sites were found.The activitation of Cbfαl was systematically tested in Hela cell by co-transfection of PCMV5-Cbfαl with 6 different Amelogenin promoter report gene.The longest promoter report gene was co-transfected into Hela, CHO, and UMR-106 cells with PCMV5-Cbfαl to compare the different roles of Cbfαl between Hela,CHO,and UMR-106 cells.RESULTS:Cbfαl served as a strong transcriptional activator on Amelogenin promoter in Hela cell,and this activating degree kept the same lever on different deleted promoter.In CHO and UMR-106 cells,Cbfαl showed a weak activation.CONCLUSION:In Hela cells, according to the promoting ways of Cbfαl acting on Amelogenin promoter,the potential Cbfαl binding site in Amelogenin promoter seemed located in the first intron of Amelogenin gene.In term of the weak activation of Cbfαl in CHO and UMR-106 cells,It proved that this activation is epithelium-specific.
出处
《牙体牙髓牙周病学杂志》
CAS
2005年第4期185-188,共4页
Chinese Journal of Conservative Dentistry
