Mima Kazuko, Donai Hitomi, Yamauchi Takashi
Department of Biochemistry, Faculty of Pharmaceutical Sciences, Tokushima University. Shomachi 1, Tokushima 770-8505. Japan.
Biol Proced Online. 2002 Apr 12;3:79-90. doi: 10.1251/bpo26.
The promoter activity of the rat Ca(2+)/calmodulin-dependent protein kinase II gene was analyzed using the luciferase reporter gene in neuronal and non-neuronal cell lines. Neuronal cell type-specific promoter activity was found in the 5'-flanking region of alpha and beta isoform genes of the kinase. Silencer elements were also found further upstream of promoter regions. A brain-specific protein bound to the DNA sequence of the 5'-flanking region of the gene was found by gel mobility shift analysis in the nuclear extract of the rat brain, including the cerebellum, forebrain, and brainstem, but not in that of non-neuronal tissues, including liver, kidney and spleen. The luciferase expression system and gel shift analysis can be used as an additional and better index by which to monitor gene expression in most cell types.
利用荧光素酶报告基因,在神经元和非神经元细胞系中分析了大鼠钙/钙调蛋白依赖性蛋白激酶II基因的启动子活性。在该激酶α和β亚型基因的5'侧翼区域发现了神经元细胞类型特异性启动子活性。在启动子区域的更上游也发现了沉默子元件。通过凝胶迁移率变动分析,在大鼠脑(包括小脑、前脑和脑干)的核提取物中发现一种脑特异性蛋白与该基因5'侧翼区域的DNA序列结合,但在包括肝、肾和脾在内的非神经元组织的核提取物中未发现。荧光素酶表达系统和凝胶迁移率变动分析可作为监测大多数细胞类型中基因表达的额外且更好的指标。