Bram R J, Kornberg R D
Proc Natl Acad Sci U S A. 1985 Jan;82(1):43-7. doi: 10.1073/pnas.82.1.43.
A binding activity specific for the upstream activating sequence of the GAL1-GAL10 promoter of Saccharomyces cerevisiae has been purified 220-fold on the basis of a nitrocellulose filter-binding assay. The binding activity is enriched in a nuclear preparation and is likely to be the GAL4 gene product. DNase I-protection mapping patterns reveal binding to two 30-base-pair regions at the boundaries of the sequence. A nearly identical mapping pattern is obtained with the coordinately regulated GAL7 promoter. The four 30-base-pair regions of binding in the two promoters are closely homologous, with a core consensus sequence of C-G-CG-TG-C-A-A-C-A-G-T-G-C-T-C-C-G-A-A- GC-G-A-T. A synthetic oligonucleotide with such a sequence competes with the upstream activating sequence in the binding reaction.
基于硝酸纤维素滤膜结合试验，对酿酒酵母GAL1 - GAL10启动子上游激活序列具有特异性的一种结合活性已被纯化了220倍。这种结合活性在细胞核提取物中得到富集，并且很可能是GAL4基因的产物。DNase I保护图谱分析模式显示，它与该序列边界处的两个30碱基对区域结合。对于协同调控的GAL7启动子，也获得了几乎相同的图谱分析模式。这两个启动子中四个30碱基对的结合区域高度同源，其核心共有序列为C - G - CG - TG - C - A - A - C - A - G - T - G - C - T - C - C - G - A - A - GC - G - A - T。具有这种序列的合成寡核苷酸在结合反应中与上游激活序列竞争。
