<br>评估离子强度对 PNA:DNA 双链体形成动力学的影响,RSC Chemical Biology

当前位置： X-MOL 学术 › RSC Chem. Biol. › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

评估离子强度对 PNA:DNA 双链体形成动力学的影响
RSC Chemical Biology ( IF 4.2 ) Pub Date : 2021-5-21 , DOI: 10.1039/d1cb00025j
Colin S Swenson ₁ , Hershel H Lackey ₂ , Eric J Reece ₂ , Joel M Harris ₂ , Jennifer M Heemstra ₁ , Eric M Peterson ₂

Affiliation

肽核酸（PNA）是一种独特的合成核酸类似物，已被用于许多生物应用。这些应用依赖于 PNA 提供的强大的 Franklin-Watson-Crick 碱基配对，特别是在离子强度较低的情况下。然而，我们对 PNA:DNA 杂交动力学与离子强度之间关系的理解并不完整。在这里，我们使用全内反射荧光成像在单分子分辨率下测量了 PNA 与 DNA 在一系列离子强度和温度下的结合和解离动力学。与 DNA:DNA 双链体不同，PNA:DNA 双链体在较低离子强度下更稳定，我们证明这是由于较高的结合率。虽然 PNA:DNA 双链体的解离速率对离子强度基本上不敏感，但它明显低于具有相同数量和序列的碱基配对相互作用的 DNA:DNA 双链体的解离速率。 PNA:DNA 动力学速率常数的温度依赖性表明对双链体解离以及在较小程度上对双链体形成存在显着的焓障碍。这项对 PNA:DNA 杂交动力学的研究为更好地理解和设计 PNA 序列以供未来应用提供了一个框架。

"点击查看英文标题和摘要"

Evaluating the effect of ionic strength on PNA:DNA duplex formation kinetics

Peptide nucleic acid (PNA) is a unique synthetic nucleic acid analog that has been adopted for use in many biological applications. These applications rely upon the robust Franklin–Watson–Crick base pairing provided by PNA, particularly at lower ionic strengths. However, our understanding of the relationship between the kinetics of PNA:DNA hybridization and ionic strength is incomplete. Here we measured the kinetics of association and dissociation of PNA with DNA across a range of ionic strengths and temperatures at single-molecule resolution using total internal reflection fluorescence imaging. Unlike DNA:DNA duplexes, PNA:DNA duplexes are more stable at lower ionic strength, and we demonstrate that this is due to a higher association rate. While the dissociation rate of PNA:DNA duplexes is largely insensitive to ionic strength, it is significantly lower than that of DNA:DNA duplexes having the same number and sequence of base pairing interactions. The temperature dependence of PNA:DNA kinetic rate constants indicate a significant enthalpy barrier to duplex dissociation, and to a lesser extent, duplex formation. This investigation into the kinetics of PNA:DNA hybridization provides a framework towards better understanding and design of PNA sequences for future applications.

更新日期：2021-05-28

点击分享查看原文

点击收藏

公开下载

阅读更多本刊新发论文