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长均匀G4-DNA纳米线的高效制备方法及其性质

Efficient procedure of preparation and properties of long uniform G4-DNA nanowires.

作者信息

Borovok Natalia, Molotsky Tatiana, Ghabboun Jamal, Porath Danny, Kotlyar Alexander

机构信息

Department of Biochemistry, George S. Wise Faculty of Life Sciences, Tel Aviv University, Aviv 69978, Israel.

出版信息

Anal Biochem. 2008 Mar 1;374(1):71-8. doi: 10.1016/j.ab.2007.10.017. Epub 2007 Oct 22.

Abstract

Here we describe a novel and efficient procedure for preparation of long uniform G4-DNA wires. The procedure includes (i) enzymatic synthesis of double-stranded DNA molecules consisting of long (up to 10,000 bases), continuous G strands and chains of complementary (dC)20-oligonucleotides, poly(dG)-n(dC)20; (ii) size exclusion HPLC separation of the G strands from the (dC)20 oligonucleotides in 0.1M NaOH; and (iii) folding of the purified G strands into G4-DNA structures by lowering the pH to 7.0. We show by atomic force microscopy (AFM) that the preparation procedure yielded G4-DNA wires with a uniform morphology and a narrow length distribution. The correlation between the total amount of nucleotides in the G strands and the contour length of the G4-DNA molecules estimated by AFM suggests monomolecular folding of the G strands into quadruplex structures. The folding takes place either in the presence or in the absence of stabilizing ions (K+ or Na+). The addition of these cations leads to a dramatic change in the circular dichroism spectrum of the G4-DNA.

摘要

在此,我们描述了一种制备长且均匀的G4-DNA链的新颖且高效的方法。该方法包括:(i)酶促合成由长(长达10,000个碱基)的连续G链和互补的(dC)20寡核苷酸链组成的双链DNA分子,即聚(dG)-n(dC)20;(ii)在0.1M NaOH中通过尺寸排阻高效液相色谱法从(dC)20寡核苷酸中分离G链;(iii)通过将pH值降至7.0,将纯化的G链折叠成G4-DNA结构。我们通过原子力显微镜(AFM)表明,该制备方法产生了具有均匀形态和窄长度分布的G4-DNA链。G链中核苷酸的总量与通过AFM估计的G4-DNA分子的轮廓长度之间的相关性表明,G链单分子折叠成四链体结构。折叠过程在有或没有稳定离子(K +或Na +)的情况下均可发生。添加这些阳离子会导致G4-DNA的圆二色光谱发生显著变化。

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