Effect of sodium ions on RNA duplex stability.

Effect of sodium ions on RNA duplex stability.
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DOI:
10.1021/bi4008275
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发表时间:
2013-10-22
期刊:
影响因子:
2.9
通讯作者:
Znosko, Brent M.
Znosko, Brent M.
中科院分区:
生物学3区
文献类型:
--
作者:
Chen, Zexiang;Znosko, Brent M.

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RNA 光学熔解实验的标准钠浓度为 1.021 M。根据序列预测 Tm、ΔG°37 和二级结构的算法通常依赖于在 1.021 M 钠中进行的光学熔解实验得出的参数。生理单价阳离子浓度远低于 1.021 M。事实上,许多分子生物学技术需要含有 1.021 M 以外的单价阳离子浓度的缓冲液。当单价阳离子浓度不是 1.021 M 时,基于 1.021 M Na+ 参数的预测可能不准确。在这里,我们报告了一组 18 个 RNA 双链体的光学熔解实验的热力学数据,每个双链体在较宽的钠离子浓度范围内熔解(71、121、221 和 621 mM)。使用该数据和先前发布的在 1.021 M Na+ 中熔化的相同序列的数据,我们报告了 Tm 和 ΔG°37 校正因子,以将标准 1.021 M Na+ RNA 参数缩放到其他钠离子浓度。推荐的 Tm 校正因子(eq 21)预测熔化温度在 0.7 °C 以内,推荐的 ΔG°37 校正因子(eq 26)预测自由能在 0.14 kcal/mol 以内。这些校正因子可以纳入预测算法中,根据序列预测 RNA 二级结构,并提供 RNA 双链体的 Tm 和 ΔG°37 值。
The standard sodium concentration for RNA optical melting experiments is 1.021 M. Algorithms that predict Tm, ΔG°37, and secondary structure from sequence generally rely on parameters derived from optical melting experiments performed in 1.021 M sodium. Physiological monovalent cation concentrations are much lower than 1.021 M. In fact, many molecular biology techniques require buffers containing monovalent cation concentrations other than 1.021 M. Predictions based on the 1.021 M Na+ parameters may not be accurate when the monovalent cation concentration is not 1.021 M. Here, we report thermodynamic data from optical melting experiments for a set of 18 RNA duplexes, each melted in a wide range of sodium ion concentrations (71, 121, 221, and 621 mM). Using this data and previously published data for the same sequences melted in 1.021 M Na+, we report Tm and ΔG°37 correction factors to scale the standard 1.021 M Na+ RNA parameters to other sodium ion concentrations. The recommended Tm correction factor (eq 21) predicts the melting temperature within 0.7 °C, and the recommended ΔG°37 correction factor (eq 26) predicts the free energy within 0.14 kcal/mol. These correction factors can be incorporated into prediction algorithms that predict RNA secondary structure from sequence and provide Tm and ΔG°37 values for RNA duplexes.
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