SHAPE analysis of small RNAs and riboswitches.
SHAPE analysis of small RNAs and riboswitches.
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DOI：
10.1016/b978-0-12-801122-5.00008-8
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发表时间：
2014
影响因子：
--
通讯作者：
Weeks KM
中科院分区：
文献类型：
--
作者：
Rice GM;Busan S;Karabiber F;Favorov OV;Weeks KM
我们描述了通过SHAPE化学探针的RNA的结构分析。用1-甲基-7-硝基靛红酸酐（1 M7）（一种检测局部核苷酸柔性的试剂）和N-甲基靛红酸酐（匪IA）和1-甲基-6-硝基靛红酸酐（1 M6）（一起检测高阶和非典型相互作用的试剂）处理RNA。在逆转录酶介导的引物延伸过程中，化学加合物被检测为终止物。探测信息可用于推断构象变化和配体结合，并开发高度精确的RNA二级结构模型。
We describe structural analysis of RNAs by SHAPE chemical probing. RNAs are treated with 1-methyl-7-nitroisatoic anhydride (1M7), a reagent that detects local nucleotides flexibility, and N-methylisatoic anhydride (NMIA) and 1-methyl-6-nitroisatoic anhydride (1M6), reagents which together detect higher-order and non-canonical interactions. Chemical adducts are detected as stops during reverse transcriptase-mediated primer extension. Probing information can be used to infer conformational changes and ligand binding, and to develop highly accurate models of RNA secondary structures.