Dep. of Genetics, Harvard Medical School, Boston, MA 02115, USA.
Department of Chemical-Engineering, Ben-Gurion University of the Negev, Beer Sheva 84105, Israel.
Soft Matter. 2018 Mar 28;14(12):2219-2226. doi: 10.1039/c7sm01328k. Epub 2018 Feb 16.
We utilized single-molecule tethered particle motion (TPM) tracking, optimized for studying the behavior of short (0.922 μm) dsDNA molecules under shear flow conditions, in the proximity of a wall (surface). These experiments track the individual trajectories through a gold nanobead (40 nm in radius), attached to the loose end of the DNA molecules. Under such circumstances, local interactions with the wall become more pronounced, manifested through hydrodynamic interactions. To elucidate the mechanical mechanism that affects the statistics of the molecular trajectories of the tethered molecules, we estimate the resting diffusion coefficient of our system. Using this value and our measured data, we calculate the orthogonal distance of the extended DNA molecules from the surface. This calculation considers the hydrodynamic drag effect that emerges from the proximity of the molecule to the surface, using the Faxén correction factors. Our finding enables the construction of a scenario according to which the tension along the chain builds up with the applied shear force, driving the loose end of the DNA molecule away from the wall. With the extension from the wall, the characteristic times of the system decrease by three orders of magnitude, while the drag coefficients decay to a plateau value that indicates that the molecule still experiences hydrodynamic effects due to its proximity to the wall.
我们利用单分子系链粒子运动(tethered particle motion,TPM)追踪技术,该技术经过优化,可用于在壁面(表面)附近研究短(0.922 μm)dsDNA 分子在切变流条件下的行为。这些实验通过附着在 DNA 分子松散端的金纳米珠(半径 40nm)追踪单个轨迹。在这种情况下,与壁面的局部相互作用更加明显,表现为流体动力学相互作用。为了阐明影响系链分子分子轨迹统计的力学机制,我们估计了系统的静止扩散系数。使用该值和我们测量的数据,我们计算了伸展 DNA 分子到表面的正交距离。该计算考虑了由于分子接近表面而产生的流体动力阻力效应,并使用 Faxén 修正因子。我们的发现使我们能够构建一个情景,根据该情景,链上的张力随着施加的剪切力而增加,从而将 DNA 分子的松散端从壁面拉开。随着与壁面的延伸,系统的特征时间减少了三个数量级,而阻力系数衰减到一个平台值,表明分子由于靠近壁面仍然受到流体动力效应的影响。