Fainsod A, Bogarad L D, Ruusala T, Lubin M, Crothers D M, Ruddle F H
Proc Natl Acad Sci U S A. 1986 Dec;83(24):9532-6. doi: 10.1073/pnas.83.24.9532.
Nuclear protein extracts from day 12.5 mouse embryos were used to study protein binding to DNA sequences 5' of the Hox 1.5 homeo box. Embryos of this developmental stage are known to express this gene. DNA binding protein blotting and retardation gel techniques show that murine embryonic nuclear proteins specifically bind a 753-base pair (bp) DNA fragment from the region upstream of the Hox 1.5 homeo box. A fusion protein containing the Hox 1.5 homeo domain constructed in lambda gt11 also binds the same 753-bp DNA fragment. Specific binding of the fusion protein to the upstream DNA fragment shows that the homeo box contains the sequences required for specific protein-DNA interactions, and the 753-bp fragment contains a homeo domain binding site. These results support the hypothesis that murine homeo boxes are DNA binding domains of proteins involved in the regulation of embryonic development.
利用来自12.5天小鼠胚胎的核蛋白提取物，研究蛋白质与Hox 1.5同源异型框5'端DNA序列的结合情况。已知处于该发育阶段的胚胎会表达此基因。DNA结合蛋白印迹法和阻滞凝胶技术表明，鼠胚胎核蛋白特异性结合来自Hox 1.5同源异型框上游区域的一个753碱基对（bp）的DNA片段。在λgt11中构建的包含Hox 1.5同源结构域的融合蛋白也能结合相同的753-bp DNA片段。融合蛋白与上游DNA片段的特异性结合表明，同源异型框包含蛋白质与DNA特异性相互作用所需的序列，且该753-bp片段含有一个同源结构域结合位点。这些结果支持了这样一种假说，即鼠同源异型框是参与胚胎发育调控的蛋白质的DNA结合结构域。
