Jack R S, Gehring W J, Brack C
Cell. 1981 May;24(2):321-31. doi: 10.1016/0092-8674(81)90322-6.
Using a nitrocellulose filter binding assay we have detected a protein component in extracts prepared from the nuclei of D. melanogaster third instar larvae which shows sequence-specific binding to a short region on a cloned D. melanogaster DNA segment containing a copy of the major heat-shock protein gene. The specificity of binding was confirmed by direct visualization of the protein DNA complexes in the electron microscope. Protein blotting experiments demonstrated that three polypeptides in the extract were capable of such sequence-specific binding. The protein binding site detected in this cloned sequence lies 800-1000 bp upstream from the presumptive transcription start.
通过硝酸纤维素滤膜结合试验,我们在黑腹果蝇三龄幼虫细胞核提取物中检测到一种蛋白质成分,该成分能与克隆的黑腹果蝇DNA片段上的一个短区域进行序列特异性结合,该片段含有一个主要热休克蛋白基因的拷贝。通过电子显微镜直接观察蛋白质 - DNA复合物,证实了结合的特异性。蛋白质印迹实验表明,提取物中的三种多肽能够进行这种序列特异性结合。在该克隆序列中检测到的蛋白质结合位点位于假定转录起始点上游800 - 1000 bp处。