Singh I S, Luo Z J, Eng A, Erlichman J
Department of Medicine, Albert Einstein College of Medicine, Bronx, New York 10461.
Biochem Biophys Res Commun. 1991 Jul 15;178(1):221-6. doi: 10.1016/0006-291x(91)91802-j.
The promoter and exon 1 of the regulatory subunit (RII beta) of type II cAMP-dependent protein kinase were isolated from a mouse genomic library. The 5'-flanking DNA lacked TATA and CAAT sites but contained GC rich regions typically found in constitutively expressed house keeping genes. Fusion gene constructs, containing RII beta 5'-flanking sequences and the bacterial CAT structural gene, were transfected into NB2a neuroblastoma cells and CHO cells. The NB2a cells expressed high levels of CAT activity. CHO cells expressed CAT activity at 5% of the level seen in the NB2a cells. Transfection of deletion constructs into both cell lines was used to define the core promoter and enhancer elements. The core promoter was situated between bp -291/-121. An enhancer element was located between bp -1426/-1018.
从小鼠基因组文库中分离出II型环磷酸腺苷依赖性蛋白激酶调节亚基（RIIβ）的启动子和外显子1。5'侧翼DNA缺乏TATA和CAAT位点，但含有在组成型表达的管家基因中常见的富含GC的区域。将含有RIIβ 5'侧翼序列和细菌CAT结构基因的融合基因构建体转染到NB2a神经母细胞瘤细胞和CHO细胞中。NB2a细胞表达高水平的CAT活性。CHO细胞表达的CAT活性为NB2a细胞中所见水平的5%。将缺失构建体转染到这两种细胞系中以确定核心启动子和增强子元件。核心启动子位于bp -291/-121之间。一个增强子元件位于bp -1426/-1018之间。
