Amin J, Mestril R, Lawson R, Klapper H, Voellmy R
Mol Cell Biol. 1985 Jan;5(1):197-203. doi: 10.1128/mcb.5.1.197-203.1985.
A hybrid gene in which the expression of an Escherichia coli beta-galactosidase gene was placed under the control of a Drosophila melanogaster 70,000-dalton heat shock protein (hsp70) gene promoter was constructed. Mutant derivatives of this hybrid gene which contained promoter sequences of different lengths were prepared, and their heat-induced expression was examined in D. melanogaster and COS-1 (African green monkey kidney) cells. Mutants with 5' nontranscribed sequences of at least 90 and up to 1,140 base pairs were expressed strongly in both cell types. Mutants with shorter 5' extensions (of at least 63 base pairs) were transcribed and translated efficiently in COS-1 but not at all in D. melanogaster cells. Thus, in contrast to the situation in COS-1 cells, the previously defined heat shock consensus sequence which is located between nucleotides 62 and 48 of the hsp70 gene 5' nontranscribed DNA segment is not sufficient for the expression of the D. melanogaster gene in homologous cells. A second consensus-like element 69 to 85 nucleotides upstream from the cap site is postulated to be also involved in the heat-induced expression of the hsp70 gene in D. melanogaster cells.
构建了一种杂种基因，其中大肠杆菌β-半乳糖苷酶基因的表达受果蝇70,000道尔顿热休克蛋白（hsp70）基因启动子的控制。制备了含有不同长度启动子序列的该杂种基因的突变衍生物，并在果蝇和COS-1（非洲绿猴肾）细胞中检测了它们的热诱导表达。具有至少90至1140个碱基对的5'非转录序列的突变体在两种细胞类型中均强烈表达。具有较短5'延伸（至少63个碱基对）的突变体在COS-1细胞中能有效转录和翻译，但在果蝇细胞中则完全不表达。因此，与COS-1细胞中的情况相反，位于hsp70基因5'非转录DNA区段核苷酸62至48之间的先前定义的热休克共有序列不足以使果蝇基因在同源细胞中表达。据推测，帽位点上游69至85个核苷酸处的第二个类似共有序列元件也参与了果蝇细胞中hsp70基因的热诱导表达。
