Effects of Na+ and Mg2+ on the structures of supercoiled DNAs: comparison of simulations with experiments

Recent cryo-electron microscopy (cryo-EM) results suggest that sufficient NaCl concentration (> or approximately 0.1 M) and superhelix density (> or approximately-0.05) cause circular DNAs to adopt highly extended, tightly interwound configurations, in which the strands are laterally contiguous along almost their entire length. Millimolar levels of MgCl2 reportedly act synergistically with NaCl to produce similar conformations. However, Monte Carlo simulations with purely repulsive interduplex forces failed to reproduce such structures. In the present work, solution measurements of particular physical properties were performed both to characterize the effects of Na+ and Mg2+ on DNA structure and to provide quantitative tests of Monte Carlo simulations of circular DNAs. Supercoiled p30 delta DNAs in 10 mM Tris plus 0, 0.122, and 0.1 M NaCl, and 0.1 M NaCl plus 4 mM Mg2+ were examined by static and dynamic light scattering (LS and DLS), time-resolved fluorescence polarization anisotropy (FPA) of intercalated ethidium, and circular dichroism (CD) spectroscopy. Upon addition of 0.122 M NaCl, the radius of gyration (Rg) decreased substantially, which indicates that p30 delta adopts a more compact structure. This contradicts the cryo-EM studies, where molecular extension and Rg both increase upon adding 0.1 M NaCl. In 0.1 M NaCl, the torsion constant measured by FPA is practically invariant to superhelix density, and the plateau diffusion coefficient at large scattering vector (Dplat) is likewise nearly the same at both relaxed and native superhelix densities. Such invariance is difficult to reconcile with any transition from relaxed circles to tightly interwound structures with laterally contiguous strands. Metropolis Monte Carlo simulations were performed to generate canonically distributed sets of structures, from which average Do values and scattered intensity ratios, [symbol: see text]I (zero) [symbol: see text]/[symbol: see text] l(k) [symbol: see text], were calculated. Agreement between simulations and experiments in regard to [symbol: see text] I(O) [symbol: see text] /[symbol: see text] I(k) [symbol: see text], D(zero) and the supercoiling free energy, delta Gsc (delta l), is remarkably good for the most extensively studied p30 delta samples. The simulated structures exhibit no sign of very tight interwinding with extensive lateral contacts, but instead exhibit most probable superhelix diameters of 85 to 90 A. When 4 mM Mg2+ was added to native supercoiled p30 delta in 0.1 M NaCl, Rg decreased, D(zero) increased, and the longest internal relaxation rate (1/tau 2(zero)) increased, all of which indicate a further overall contraction of the molecular envelope. The torsion constant exhibited a slight increase that is hardly statistically significant. In this case, agreement between the simulations and experiments was only semi-quantitative for most samples investigated, although the predicted contraction was exhibited by all five samples of p30 delta and one of pBR322 DNA. The simulated structures in 0.1 M NaCl plus 4 mM Mg2+ again showed no sign of extensive lateral contacts. A plausible explanation is proposed for the highly extended, tightly interwound structures seen in cryo-EM, and explicitly tested by Monte Carlo simulations of a 1000 bp circular DNA at +25 and -50 degrees C. Structures identical to those seen in cryo-EM are in fact the equilibrium structures in the simulations at -50 degrees C, and the estimated time for equilibration (2.3 x 10(-6) second) is much smaller than the estimated time for vitrification (1 x 10(-4) second).     

Racial aspects of blood pressure in children and adolescents

I have discussed a model of the psyche comprising two different modes of thinking, one non-psychotic and the other psychotic. I have related these modes of thinking to our modern myth of Jekyll and Hyde, the study of which could in my opinion give us some insight into their nature. In my view a non-psychotic state of mind belongs to a person who has a history, with particular parents, a particular development, particular conflicts, and operates in the depressive position. A psychotic state of mind belongs to a person who lives in a still and timeless present, with no origin, no development and no conflict, and operates in the paranoid-schizoid position. On the basis of this model I have subsequently described the life history of a psychotic patient and an analytic session in detail, showing how psychotic and non-psychotic states alternate and interact with each other within the same individual and between patient and therapist. The use of my countertransference, moving from a concrete to a symbolic position, has enabled me to make an interpretation. The result of this interpretation has been double, leading to a negative therapeutic reaction. An upsurge of psychotic fury was followed by increased patient/therapist communication, with a small movement from the paranoid-schizoid to the depressive position.     

姑山矿空心砖厂原料处理工艺线的实际应用探讨

文章结合制砖行业原料处理工艺的特点,阐述了姑山矿空心砖厂原料制备过程中所存在的主要问题及相应解决办法.通过对改造前后原料处理工艺的使用效果进行对比,改造后的原料处理工艺更适合于生产线的要求,并且在生产中取得了较好的实际应用效果.     

高层建筑结构设计特点与剪力墙设计

围绕高层建筑结构，总结了高层建筑结构设计的特点，提出了剪力墙设计的几个问题，以及高层建筑结构分析和各种体系相对应的方法。     

三相异步电动机运行指标的分析

利用电机学理论分析了影响电机起动转矩、起动电流、最大转矩和最小转矩的因素。     

Review of Daqing＇s Technologies in 10th Five-Year Plan Period

During the 10^th Five-Year Plan Period, Daqing Oil Field entered the late high water-cut period with the reserve production ratio in imbalance. The oil field development became more and more difficult owing to the inadequate substitute resources. Daqing Oil Field Company has made a series of important and new technological achievements to solve those bottlenecks. The oil field has cumulatively achieved a total of technological results, of which eight items are at the State level, 96 items at the provincial and ministerial level, and 1009 items granted with the patent rights. The oil field has developed a family of exploration and development technologies which are world-leading and self-developed with an independent IPR.     

钢中氧氮的测定方法研究

主要介绍了ELTRAON900气体分析仪测量O,N的原理,并重点探讨了分析参数的改变对钢中测量结果的影响,确定了测量钢中O,N的最佳分析条件,在该条件下测定钢中O,N的相对标准偏差RSD分别为2.65%～5.07%,0.63%～28.3%,符合测量要求.