Conwell Christine C, Vilfan Igor D, Hud Nicholas V
Parker H. Petit Institute for Bioengineering and Biosciences, School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, GA 30332-0400, USA.
Proc Natl Acad Sci U S A. 2003 Aug 5;100(16):9296-301. doi: 10.1073/pnas.1533135100. Epub 2003 Jul 18.
The process of DNA condensation into nanometer-scale particles has direct relevance to several fields, including cell biology, virology, and gene delivery for therapeutic purposes. DNA condensation has also attracted the attention of polymer physicists, as the collapse of DNA molecules from solution into well defined particles represents an exquisite example of a polymer phase transition. Here we present a quantitative study of DNA toroids formed by condensation of 3 kb DNA with hexammine cobalt (III). The presence or absence of static loops within this DNA molecule demonstrates the effect of nucleation loop size on toroid dimensions and that nucleation is principally decoupled from toroid growth. A comparison of DNA condensates formed at low ionic strength with those formed in the presence of additional salts (NaCl or MgCl2) shows that toroid thickness is a salt-dependant phenomenon. Together, these results have allowed the development of models for DNA toroid formation in which the size of the nucleation loop directly influences the diameter of the fully formed toroid, whereas solution conditions govern toroid thickness. The data presented illustrate the potential that exists for controlling DNA toroid dimensions. Furthermore, this study provides a set of data that should prove useful as a test for theoretical models of DNA condensation.
DNA凝聚成纳米级颗粒的过程与多个领域直接相关,包括细胞生物学、病毒学以及用于治疗目的的基因递送。DNA凝聚也引起了聚合物物理学家的关注,因为DNA分子从溶液中塌缩成定义明确的颗粒代表了聚合物相变的一个精妙例子。在此,我们对由3 kb DNA与六氨合钴(III)凝聚形成的DNA环进行了定量研究。该DNA分子中静态环的存在与否证明了成核环大小对环尺寸的影响,并且成核过程主要与环的生长解耦。将在低离子强度下形成的DNA凝聚物与在添加额外盐(NaCl或MgCl2)的情况下形成的凝聚物进行比较,结果表明环的厚度是一种依赖盐的现象。总之,这些结果使得能够建立DNA环形成的模型,其中成核环的大小直接影响完全形成的环的直径,而溶液条件决定环的厚度。所呈现的数据说明了控制DNA环尺寸的潜力。此外,本研究提供了一组数据,作为对DNA凝聚理论模型的检验应会很有用。