Department of Chemistry, Faculty of Pure and Applied Sciences, University of Tsukuba, Tsukuba 305-8571, Ibaraki, Japan.
Langmuir. 2022 Jul 12;38(27):8462-8468. doi: 10.1021/acs.langmuir.2c01062. Epub 2022 Jun 29.
In this study, we investigated the distribution behavior of single-stranded DNA molecules with 20 bases in silica particles (particle size: ∼30 μm) using confocal fluorescence microspectroscopy. The distribution kinetics was investigated under various conditions, such as the type of base (adenine, thymine, guanine, and cytosine), pore size of the particle (30 and 50 nm), and salt concentration (100, 200, and 500 mM), which changed the distribution behavior. At high salt concentrations, we observed sigmoidal kinetic behavior, which does not occur in the general distribution of small organic molecules but is often observed in protein aggregation and nuclear growth. An analytical model based on DNA aggregation explained the sigmoidal distribution behavior well, and this model also worked well when the number of DNA molecules involved in DNA aggregation was greater than two. The intraparticle diffusion of DNA molecules was analyzed using the pore and surface diffusion model. As a result, the intraparticle diffusion of DNA aggregates mainly occurs according to surface diffusion, and the surface diffusion coefficient has the same value ((2.4-6.7) × 10 cm s) independent of the pore size and type of base.
在这项研究中,我们使用共聚焦荧光显微镜研究了 20 个碱基的单链 DNA 分子在二氧化硅颗粒(粒径:约 30μm)中的分布行为。在不同的条件下研究了分布动力学,例如碱基类型(腺嘌呤、胸腺嘧啶、鸟嘌呤和胞嘧啶)、颗粒的孔径(30nm 和 50nm)和盐浓度(100、200 和 500mM),这些条件改变了分布行为。在高盐浓度下,我们观察到了 S 型动力学行为,这种行为在一般的小分子有机物质的分布中不会发生,但在蛋白质聚集和核生长中经常观察到。基于 DNA 聚集的分析模型很好地解释了 S 型分布行为,当参与 DNA 聚集的 DNA 分子数量大于两个时,该模型也能很好地工作。使用孔和表面扩散模型分析了 DNA 分子的颗粒内扩散。结果表明,DNA 聚集体的颗粒内扩散主要根据表面扩散进行,表面扩散系数与孔径和碱基类型无关,均具有相同的值((2.4-6.7)×10 cm s)。