Stefanovsky Victor Y, Moss Tom
Laboratory of Growth and Development, St-Patrick Research Group in Basic Oncology, Cancer Division of the Quebec University Hospital Research Centre, Quebec City, QC, Canada.
Department of Moleculaier Biology, Medical Biochemistry and Pathology, Faculty of Medicine, Laval University, Edifice St Patrick, 9 rue McMahon, Québec, QC, Canada, G1R 3S3.
Methods Mol Biol. 2015;1334:195-203. doi: 10.1007/978-1-4939-2877-4_12.
So-called architectural DNA-binding proteins such as those of the HMGB-box family induce DNA bending and kinking. However, these proteins often display only a weak sequence preference, making the analysis of their DNA-binding characteristics difficult if not impossible in a standard electrophoretic mobility shift assay (EMSA). In contrast, such proteins often bind prebent DNAs with high affinity and specificity. A synthetic cruciform DNA structure will often provide an ideal binding site for such proteins, allowing their affinities for both bent and linear DNAs to be directly and simply determined by a modified form of EMSA.
所谓的结构DNA结合蛋白，如HMGB框家族的蛋白，可诱导DNA弯曲和扭结。然而，这些蛋白通常仅表现出较弱的序列偏好性，这使得在标准的电泳迁移率变动分析（EMSA）中分析它们的DNA结合特性即便不是不可能，也是困难的。相比之下，这类蛋白通常以高亲和力和特异性结合预弯曲的DNA。合成十字形DNA结构通常会为此类蛋白提供理想的结合位点，通过改良形式的EMSA可以直接且简单地测定它们对弯曲和线性DNA的亲和力。
