Damante G, Rapoport B
Department of Medicine, Veterans' Administration Medical Center, San Francisco, CA 94121.
J Mol Biol. 1988 Mar 5;200(1):213-6. doi: 10.1016/0022-2836(88)90347-6.
We report the presence of an element in the rat c-Ha-ras 5'-flanking region that has a suppressive effect on the promoter of this gene. Promoter activity was determined using constructs of different regions of the c-Ha-ras 5'-flanking region linked to the reporter gene chloramphenicol acetyl transferase (CAT) transfected into NIH-3T3 cells. The presence of the tract 400 base-pairs upstream from the promoter region reduced promoter activity tenfold. This suppressor element was effective in either orientation and was not produced by other DNA fragments of similar length. Transfection with different amounts of plasmid suggested that a trans-acting factor in limiting amounts was acting on the suppressor region. Gel-retardation assays demonstrated that a nuclear protein was present in 3T3 cells that specifically interacted with the 400-base fragment containing the suppressor element.
我们报道了在大鼠c-Ha-ras基因5'侧翼区存在一种对该基因启动子具有抑制作用的元件。通过构建与报告基因氯霉素乙酰转移酶(CAT)相连的c-Ha-ras基因5'侧翼区不同区域的载体,转染至NIH-3T3细胞中来测定启动子活性。启动子区域上游400个碱基对的片段使启动子活性降低了10倍。该抑制元件在任何方向上均有效,且不是由其他长度相似的DNA片段产生的。用不同量的质粒进行转染表明,一种数量有限的反式作用因子作用于抑制区域。凝胶阻滞试验证明,3T3细胞中存在一种核蛋白,它能与含有抑制元件的400碱基片段特异性相互作用。