SHAPE to Probe RNA Structure and RNA-Protein Interactions In Vitro.
SHAPE to Probe RNA Structure and RNA-Protein Interactions In Vitro.
复制标题
SHAPE 在体外探测 RNA 结构和 RNA-蛋白质相互作用。
DOI：
10.1007/978-1-0716-3191-1_5
复制
发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Ghosh,Gourisankar
中科院分区：
文献类型：
--
作者：
Saha,Kaushik;Ghosh,Gourisankar
引物延伸法（SHAPE）分析的选择性2′-羟酰化作用可用于区分由碱基配对或蛋白质结合调节的核苷酸的柔性水平。在这种方法中，试剂与2′羟基反应形成加合物，然后通过逆转录反应检测。在特定核苷酸位置具有加合物的RNA分子的数量指示该核苷酸的SHAPE反应性。在这里，我们描述的方法探测的RNA在蛋白质的自由或蛋白质结合状态的体外形状。
Selective 2′hydroxylacylation analyzed byprimerextension (SHAPE) is used to distinguish between the levels of flexibility of nucleotides regulated by base pairing or protein binding. In this method, a reagent reacts with the 2′ hydroxyl group to form an adduct, which is then detected by reverse transcription reaction. The number of RNA molecules with an adduct at a specific nucleotide position indicates the SHAPE reactivity of that nucleotide. Here, we describe the method for probing the structure of an RNA in a protein-free or a protein-bound state by in vitro SHAPE.