Liu Z J, Moav B, Faras A J, Guise K S, Kapuscinski A R, Hackett P
Department of Genetics and Cell Biology, University of Minnesota, St. Paul 55108.
Gene. 1991 Dec 15;108(2):211-7. doi: 10.1016/0378-1119(91)90436-f.
The beta-actin-encoding gene (Act) in carp is regulated by several cis-acting regulatory elements including the evolutionarily conserved CC(A/T)6GG (CArG box or serum-response element) sequences positioned in the promoter region between the CAAT and TATA boxes and in the first intron. To address the roles of the two CArG boxes on gene expression, we replaced them with linker sequences. The CArG box in the proximal promoter was not required for promoter activity in tissue-cultured cells, but was required in conjunction with a second CArG box in the first intron to give full expression in transgenic embryos. Likewise, the geometry of cis-acting transcriptional elements in the proximal promoter was more important for expression of transgenic constructs in developing embryos than in tissue-cultured fibroblasts. Mobility-shift and exonuclease mapping experiments indicated that the same or similar protein factors bind around the two CArG boxes, suggesting that interactions between the promoter and the first intron are involved in Act regulation.
鲤鱼中编码β-肌动蛋白的基因（Act）受几个顺式作用调控元件的调节，这些元件包括位于CAAT盒和TATA盒之间的启动子区域以及第一个内含子中的进化保守CC(A/T)6GG（CArG盒或血清反应元件）序列。为了研究两个CArG盒对基因表达的作用，我们用连接序列替换了它们。近端启动子中的CArG盒对于组织培养细胞中的启动子活性不是必需的，但与第一个内含子中的第二个CArG盒一起对于转基因胚胎中的充分表达是必需的。同样，近端启动子中顺式作用转录元件的几何结构对于发育中的胚胎中转基因构建体的表达比在组织培养的成纤维细胞中更重要。迁移率变动和核酸外切酶作图实验表明，相同或相似的蛋白质因子结合在两个CArG盒周围，这表明启动子和第一个内含子之间的相互作用参与了Act的调控。
