Adams E C, Gurley W B
Department of Microbiology and Cell Science, University of Florida, Gainesville 32611.
Plant Mol Biol. 1994 Oct;26(1):377-92. doi: 10.1007/BF00039547.
A 16 bp site of protein binding has been identified in the promoter of the 780 gene of T-DNA. Specific DNA-protein interactions were demonstrated between a double-stranded oligonucleotide containing this element (5'-TTGAAAAATCAACGCT-3') and a protein isolated from nuclear extracts of cauliflower inflorescences. Specific bases required for this binding activity (780 binding protein; 780BP) were defined by kinetic competition studies with mutated oligonucleotides, methylation interference assays and DNAse I footprinting. 780BP binding was not competed with up to 1000-fold excess of previously characterized plant regulatory elements such as as-1, the LRE, and the ocs, G-box, and AT-rich elements. In addition, 780BP was shown to bind sequences overlapping a mammalian hormone receptor element with greater affinity than the 780 element.
已在T-DNA的780基因启动子中鉴定出一个16bp的蛋白质结合位点。在含有该元件的双链寡核苷酸(5'-TTGAAAAATCAACGCT-3')与从菜花花序核提取物中分离出的一种蛋白质之间,证实了特异性的DNA-蛋白质相互作用。通过与突变寡核苷酸的动力学竞争研究、甲基化干扰分析和DNA酶I足迹分析,确定了这种结合活性(780结合蛋白;780BP)所需的特定碱基。780BP结合不会被高达1000倍过量的先前表征的植物调控元件(如as-1、LRE以及ocs、G-box和富含AT的元件)所竞争。此外,研究表明780BP与重叠哺乳动物激素受体元件的序列结合,其亲和力高于780元件。