双链DNA微阵列原理技术和应用-遗传.PDF

HEREDITAS (Beijing) 2013 3 , 35(3): 287―306 ISSN 0253-9772 综 述 DOI: 10.3724/SP.J.1005.2013.00287 双链DNA 微阵列：原理、技术和应用 潘艳, 王进科 , 210096 双链 DNA 微阵列也称为蛋白结合微阵列, 是高通量检测分析 DNA 结合蛋白(如转录因子) 与大量DNA 分子相互作用的一种重要技术。它将大量双链DNA 分子固定在特定的固相支持物(如玻片)上, 与待测蛋白相互 作用, 用以确定转录因子的DNA 结合亲和性、特异性及序列偏好性。近年来, 该技术在快速表征大量转录因子 的DNA 特异性、绘制转录因子DNA 结合谱、鉴定转录因子DNA 结合位点和靶基因、识别转录因子家族内不 同成员及其二聚体的细微DNA 结合差异、考察辅助因子对转录因子 DNA 结合特异性影响等方面展现了其重 要的应用价值。文章对双链DNA 微阵列的原理、技术及应用进行了综述。 双链DNA 微阵列; 转录因子 Double-stranded DNA microarray: principal, techniques and appli- cations PAN Yan, WANG Jin-Ke The State Key Laboratory of Bioelectronics, Southeast University, Nanjing 210096, China Abstract: Double-stranded DNA (dsDNA) microarray, also known as protein binding microarray (PBM), is an impor- tant technique that can be used to assay the interaction of DNA-binding protein (such as transcription factor, TF) with vast amount of DNA molecules in high-throughput format. This technique immobilizes large amount of various dsDNA mole- cules on the surface of a solid support (such as glass slide) for detecting the binding interaction of a DNA-binding protein with all of the immobilized dsDNA molecules, and thus determining the DNA-binding affinity, specificity and preference of TFs. In recent years, this technique has demonstrated its valuable applications in several aspects, including rapidly charac- terizing DNA-binding specificity of large number of TFs, building DNA-binding profiles of TFs, identifying DNA-binding sites and target genes of TFs, discriminating the subtle DNA-binding preferences of members and their dimmers of a TF family, and examining the effects of a cofactor on the DNA-binding specificity of TFs. This paper reviews the principal, techniques, and applicati