Thomas Jason M, Yu Hua-Zhong, Sen Dipankar
Department of Chemistry, Simon Fraser University, Burnaby, BC, Canada.
Methods Mol Biol. 2014;1103:267-76. doi: 10.1007/978-1-62703-730-3_19.
Aptamers have proven to be very useful as high-affinity and -specificity molecular recognition elements in analytical sensors of various forms. Herein, we describe a general process for creating an aptamer-based sensor that functions as an analyte-responsive, nano-sized, electronic switch. These sensors can provide an electrochemical readout, by switching through-DNA charge transfer across a DNA three-way junction from "off" to "on" in response to the binding of a target ligand to the sensor's aptamer domain. We detail the general design principles for such sensors, as well as the biochemical charge transfer assays used to identify functional sensors. In these gel electrophoresis-based assays, analyte-responsive conductivity switching is detected conveniently through biochemical experiments that characterize oxidative DNA damage patterns in sequencing PAGE gels.
适配体已被证明在各种形式的分析传感器中作为高亲和力和高特异性的分子识别元件非常有用。在此，我们描述了一种创建基于适配体的传感器的通用过程，该传感器用作分析物响应型纳米级电子开关。这些传感器可以通过响应目标配体与传感器适配体结构域的结合，使通过DNA三向连接的DNA电荷转移从“关闭”切换到“开启”来提供电化学读出。我们详细阐述了此类传感器的一般设计原则，以及用于识别功能传感器的生化电荷转移测定法。在这些基于凝胶电泳的测定法中，通过表征测序PAGE凝胶中氧化DNA损伤模式的生化实验，可以方便地检测分析物响应型电导率切换。
