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本文报告芜菁黄花叶病毒(turnip yellow mosaic virus, TYMV)RNA3′末端tRNA-样结构的热解除折迭研究结果。酶和化学探针已广泛用于RNA的二级和三级结构的测定,通过这些探针也能获得RNA分子内各种结构成分的稳定性和熔解行为的信息。在不同温度条件下对3′末端特异性标记的112个核苷酸残基的TYMV-RNA片段,用二乙基焦碳酸酯(DEP)、重亚硫酸钠,RNase T1和核酸酶S等能区分双链和单链区域。T1和S特别适宜于RNA单链的定位。T1不仅可以在各种温度(0～80℃)条件下,而且也可在不同缓冲液中进行探查。DEP用于硷基对或堆砌的腺苷残基,重亚硫酸钠能使胞苷转变成尿苷,对空间关系的探查特别有用。tRNA-样结构的大部分核苷酸均可用这些方法进行分析,它们的二级或三级结构的熔解行为与个别核苷酸或硷基对各种探针的依赖温度的易感性不同有关。
This article reports the results of pyrogenic deconvolution studies of the tRNA-like structure at the 3 ’end of turnip yellow mosaic virus (TYMV) RNA. Enzymes and chemical probes have been used extensively for the determination of the secondary and tertiary structures of RNA. These probes can also obtain information on the stability and melting behavior of various structural components in RNA molecules. The TYMV-RNA fragment of 112 nucleotides specifically labeled at the 3 ’end under different temperature conditions can be distinguished by using diethyl pyrocarbonate (DEP), sodium bisulfite, RNase T1 and nuclease S etc. Chain and single stranded region. T1 and S are particularly suitable for the localization of RNA single strands. T1 not only in a variety of temperature (0 ~ 80 ℃) conditions, but also in different buffers for exploration. DEP is used for base-pairing or stacking adenosine residues. Sodium metabisulfite converts cytidine to uridine and is particularly useful for exploring spatial relationships. Most of the nucleotides of a tRNA-like structure can be analyzed by these methods. The melting behavior of their secondary or tertiary structure is related to the temperature-dependent susceptibility of individual probes to individual nucleotides or bases.