Nickol J M, Felsenfeld G
Laboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, Bethesda, MD 20892.
Proc Natl Acad Sci U S A. 1988 Apr;85(8):2548-52. doi: 10.1073/pnas.85.8.2548.
An enhancer specific to erythroid cells was identified previously in the 3' flanking sequence of the chicken adult beta-globin gene and shown to act on the beta-globin promoter. This enhancer lies between the adult beta-globin gene and the embryonic epsilon-globin gene, about equidistant from the two promoters. To determine whether this enhancer acts also on the epsilon-globin promoter, we constructed plasmids containing the enhancer and either the beta- or the epsilon-globin promoter fused to the bacterial chloramphenicol acetyltransferase gene. Primary chicken erythrocytes of both primitive and definitive lineages were transfected with these plasmids. We show that the enhancer is able to stimulate expression from the epsilon-globin promoter as well as the beta-globin promoter. Levels of expression change with the developmental stage of the cell in a way that is partially consistent with the observed developmental regulation of the beta- and epsilon-globin genes in vivo. There appear to be no other enhancer elements either 5' of the epsilon-globin gene or within 6 kilobase pairs of its 3' end. Thus, the enhancer between the beta- and epsilon-globin genes apparently serves to regulate both genes.
先前在鸡成年β-珠蛋白基因的3'侧翼序列中鉴定出一种红系细胞特异性增强子,它可作用于β-珠蛋白启动子。该增强子位于成年β-珠蛋白基因和胚胎ε-珠蛋白基因之间,与两个启动子的距离大致相等。为了确定该增强子是否也作用于ε-珠蛋白启动子,我们构建了含有增强子以及与细菌氯霉素乙酰转移酶基因融合的β-或ε-珠蛋白启动子的质粒。用这些质粒转染了原始和定型谱系的原代鸡红细胞。我们发现该增强子能够刺激ε-珠蛋白启动子以及β-珠蛋白启动子的表达。表达水平随细胞的发育阶段而变化,这种变化方式与体内观察到的β-和ε-珠蛋白基因的发育调控部分一致。在ε-珠蛋白基因的5'端或其3'端6千碱基对内似乎没有其他增强子元件。因此,β-和ε-珠蛋白基因之间的增强子显然用于调控这两个基因。