Pinkert C A, Ornitz D M, Brinster R L, Palmiter R D
Laboratory of Reproductive Physiology, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, 19104.
Genes Dev. 1987 May;1(3):268-76. doi: 10.1101/gad.1.3.268.
Transgenic mice were used to locate the cis-acting DNA elements that are important for efficient, tissue-specific expression of the mouse albumin gene in the adult. Chimeric genes with up to 12 kb of mouse albumin 5'-flanking region fused to a human growth hormone (hGH) reporter gene were tested. Remarkably, a region located 8.5-10.4 kb upstream of the albumin promoter was essential for high-level expression in adult liver and the region in between -8.5 and -0.3 kb was dispensable. The far-upstream region behaved like an enhancer in that its position and orientation relative to the albumin promoter were not critical; however, it did not function well with a heterologous promoter. Two of four DNase hypersensitive sites found in the 5'-flanking region of the albumin gene map to the far-upstream and promoter regions; the others may reflect regions involved in developmental or environmental control of this gene.
转基因小鼠被用于定位顺式作用DNA元件,这些元件对于成年小鼠白蛋白基因的高效、组织特异性表达至关重要。对与人类生长激素(hGH)报告基因融合的长达12 kb的小鼠白蛋白5'侧翼区域的嵌合基因进行了测试。值得注意的是,位于白蛋白启动子上游8.5 - 10.4 kb的区域对于成年肝脏中的高水平表达至关重要,而位于-8.5和-0.3 kb之间的区域则是可有可无的。这个远上游区域的作用类似于增强子,因为它相对于白蛋白启动子的位置和方向并不关键;然而,它与异源启动子一起作用时效果不佳。在白蛋白基因5'侧翼区域发现的四个DNase超敏位点中的两个定位到远上游和启动子区域;其他位点可能反映了参与该基因发育或环境调控的区域。