邻位连接技术_一种新的高灵敏度蛋白质检测技术.pdf

ISSN 1007 7626 CN 11 3870 Q 中国生物化学与分子生物学报 Chinese Journal of Biochemistry and Molecular Biology 2008年 4 月 24( 4) : 383~ 389 技术与方法 邻位连接技术:一种新的高灵敏度蛋白质检测技术 李丹滢 1) , 周国华 1) , 2) * ( 1)中国药科大学生命科学与技术学院,南京 210009; 2) 华东医学生物技术研究所,南京 210002) 摘要 蛋白质组学( proteomics)诞生以来,高效准确的蛋白质检测技术受到越来越多的关注.最近, 一种高灵敏度的蛋白质检测技术, 邻位连接技术( proximity ligation assay, PLA)被建立. 该技术采用 核酸适体( aptamer)或单 多克隆抗体 核酸复合物作为邻位连接探针( proximity probes) . 当一对邻位 探针同时识别同一个目标蛋白分子时,它们将在空间位置上相互临近,通过连接反应形成一段可扩 增的 DNA标签序列,该标签序列能够反映待测蛋白的种类及浓度. 该技术将对蛋白质的检测转变 为对 DNA核酸序列的检测,实现了特殊蛋白质的检测,定量及定位.文章从该方法的产生背景,发 展过程,原理以及探针制备等方面对该方法进行了系统的介绍, 列举了该方法的几种重要应用,并 对该方法在蛋白质组学研究领域的应用前景进行了展望. 关键词 邻位连接技术; 蛋白质组学; 蛋白质检测 中图分类号 Q51 Proximity Ligation Assay : A New and Highly Sensitive Protein Detection Method LI Dan Ying 1) , ZHOU Guo Hua 1) , 2) * ( 1) College of Lif e Science and Technology, China Pharmaceutical University, Nanjing 210009, China; 2) Huadong Research Institute f or Medicine and Biotechnics, Nanj ing 210002, China ) Abstract Efficient and precise techniques for protein detection are crucial in proteomics studies. Recently, proximity ligation assay ( PLA ) , a new protein detection technique in proteomics, was developed, and attracting more and more researchers in the field. The proximity probes, aptamers or complexes of mono polyclonal ant ibody with oligonucleotide, are designed to bind pairwise to target proteins and be able to give rise to an amplifiable DNA sequence tag by ligation when brought in proximity, which reflect the identit ies and amounts of the target proteins. This approach employs the analysis of DNA sequences instead of the direct detection of specific proteins, thus achieve high - sensitivity for the detection, quantificat ion, and even localization of targeted proteins. The basic principle and the potent ial applications of PLA, as well as some technical issues, such as the preparation of the proximity probes, will be discussed in this review. Key words proximity ligation assay; proteomics; protein detection 收稿日期