Knoll B J, Zarucki-Schulz T, Dean D C, O'Malley B W
Nucleic Acids Res. 1983 Oct 11;11(19):6733-54. doi: 10.1093/nar/11.19.6733.
In order to study the initiation of transcription from the ovalbumin gene promoter, we constructed a hybrid gene (ovalglobin) in which 753 bps of ovalbumin gene 5'-flanking sequence were joined to the chicken adult beta-globin gene. When transfected into HeLa S3 cells, ovalglobin gene transcription initiated at the ovalbumin gene cap site, as measured by S1 nuclease and primer extension analysis. Deletion of 5'-flanking sequences to position -95 had little effect on transcription; deletion to -77 reduced transcription to about 20% of the wild type level and deletion to -48 reduced the level to about 2%. A deletion to -24, removing the sequence TATATAT, abolished transcription entirely. Hormonal regulation of the ovalglobin gene was observed when primary oviduct cells were used as recipients for DNA transfection. Under these conditions, addition of progesterone increased the level of ovalglobin transcripts to more than 10 times the uninduced level.
为了研究卵清蛋白基因启动子的转录起始，我们构建了一个杂种基因（卵珠蛋白），其中753个碱基对的卵清蛋白基因5'侧翼序列与鸡成体β-珠蛋白基因相连。当转染到HeLa S3细胞中时，通过S1核酸酶和引物延伸分析测定，卵珠蛋白基因转录在卵清蛋白基因帽位点起始。将5'侧翼序列缺失至-95位对转录影响不大；缺失至-77位使转录降至野生型水平的约20%，缺失至-48位使水平降至约2%。缺失至-24位，去除序列TATATAT，完全消除了转录。当将原代输卵管细胞用作DNA转染的受体时，观察到了卵珠蛋白基因的激素调节。在这些条件下，添加孕酮使卵珠蛋白转录本水平增加到未诱导水平的10倍以上。
