Peattie D A, Gilbert W
Proc Natl Acad Sci U S A. 1980 Aug;77(8):4679-82. doi: 10.1073/pnas.77.8.4679.
Three chemical reactions can probe the secondary and tertiary interactions of RNA molecules in solution. Dimethyl sulfate monitors the N-7 of guanosines and senses tertiary interactions there, diethyl pyrocarbonate detects stacking of adenosines, and an alternate dimethyl sulfate reaction examines the N-3 of cytidines and thus probes base pairing. The reactions work between 0 degrees C and 90 degrees C and at pH 4.5--8.5 in a variety of buffers. As an example we follow the progressive denaturation of yeast tRNAPhe terminally labeled with 32P as the tertiary and secondary structures sequentially melt out. A single autoradiograph of a terminally labeled molecule locates regions of higher-order structure and identifies the bases involved.
三种化学反应可用于探测溶液中RNA分子的二级和三级相互作用。硫酸二甲酯监测鸟苷的N-7，并感知那里的三级相互作用；焦碳酸二乙酯检测腺苷的堆积；另一种硫酸二甲酯反应检测胞苷的N-3，从而探测碱基配对。这些反应在0摄氏度至90摄氏度之间、pH值为4.5至8.5的各种缓冲液中均可进行。例如，我们追踪用32P末端标记的酵母苯丙氨酸tRNA随着三级和二级结构依次解链而逐渐变性的过程。对末端标记分子的一张放射自显影片可定位高阶结构区域并识别其中涉及的碱基。
