Di Primo Carmelo, Dausse Eric, Toulmé Jean-Jacques
ARNA laboratory, University of Bordeaux, F-33000 Bordeaux, France.
Methods Mol Biol. 2011;764:279-300. doi: 10.1007/978-1-61779-188-8_19.
Instruments based on the surface plasmon resonance (SPR) principle allow label-free detection of interactions between targets immobilized at a solid-liquid interface and partners in solution. This method is well suited to determine the kinetic parameters, the equilibrium constant and the stoichiometry of a reaction. Aptamers are ligands identified from random libraries of RNA, DNA or chemically modified oligonucleotides by in vitro selection (SELEX). Aptamers can be raised against a great variety of targets (small molecules, proteins, nucleic acids, cells, viruses, bacteria). SPR is routinely used in our laboratory for the analysis of RNA aptamer-RNA target complexes. To illustrate SPR investigation of such complexes, we describe here methods that were successfully used to monitor the interaction between the trans-activating responsive element of HIV-1 and an RNA aptamer.
基于表面等离子体共振（SPR）原理的仪器能够对固定在固液界面的靶标与溶液中的配体之间的相互作用进行无标记检测。该方法非常适合用于确定反应的动力学参数、平衡常数和化学计量。适体是通过体外筛选（SELEX）从RNA、DNA或化学修饰的寡核苷酸随机文库中鉴定出的配体。适体可以针对多种靶标产生（小分子、蛋白质、核酸、细胞、病毒、细菌）。SPR在我们实验室中常用于分析RNA适体-RNA靶标复合物。为了说明对这类复合物的SPR研究，我们在此描述了成功用于监测HIV-1反式激活应答元件与RNA适体之间相互作用的方法。
