聚合物支持的RNA合成及其在测试双链稳定性最近邻模型中的应用。

Polymer-supported RNA synthesis and its application to test the nearest-neighbor model for duplex stability.

作者信息

Kierzek R, Caruthers M H, Longfellow C E, Swinton D, Turner D H, Freier S M

出版信息

Biochemistry. 1986 Dec 2;25(24):7840-6. doi: 10.1021/bi00372a009.

Abstract

A solid-phase method using a phosphoramidite approach is described for synthesis of oligoribonucleotides. The method was used to synthesize pairs of oligomers with identical nearest neighbors but different sequences. Comparison of thermodynamic parameters for these pairs provides a test of the nearest-neighbor hypothesis for prediction of helix stability. In general, pairs of sequences with identical nearest neighbors have enthalpy and entropy changes for helix formation that differ by 8% on average, delta Go37 that differ by 6% on average, and melting temperatures within 0-5 degrees C of each other. These limits are typical of the accuracy that should be expected from nearest-neighbor predictions of RNA helix stability. UCAUGA and UGAUCA have the same nearest neighbors but melting temperatures that differ by 7 degrees C. This suggests some sequences will not be approximated well by the nearest-neighbor model.

摘要

描述了一种使用亚磷酰胺方法的固相法来合成寡核糖核苷酸。该方法用于合成具有相同最近邻但不同序列的寡聚物对。对这些寡聚物对的热力学参数进行比较，为预测螺旋稳定性的最近邻假设提供了检验。一般来说，具有相同最近邻的序列对在形成螺旋时的焓变和熵变平均相差8%，37℃时的ΔG₀平均相差6%，且解链温度彼此相差0 - 5℃。这些限度是RNA螺旋稳定性最近邻预测所应期望的典型精度。UCAUGA和UGAUCA具有相同的最近邻，但解链温度相差7℃。这表明某些序列不能被最近邻模型很好地近似。