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

被引:40
作者
Borovok, Natalia [2 ]
Molotsky, Tatiana [2 ]
Ghabboun, Jamal [1 ]
Porath, Danny [1 ,3 ]
Kotlyar, Alexander [2 ,4 ]
机构
[1] Hebrew Univ Jerusalem, Dept Phys Chem, IL-91904 Jerusalem, Israel
[2] Tel Aviv Univ, George S Wise Fac Life Sci, Dept Biochem, IL-69978 Tel Aviv, Israel
[3] Hebrew Univ Jerusalem, Ctr Nanosci & Nanotechnol, IL-91904 Jerusalem, Israel
[4] Tel Aviv Univ, Nanotechnol Ctr, IL-69978 Tel Aviv, Israel
基金
欧盟地平线“2020”;
关键词
G4-DNA nanowires; enzymatic synthesis; folding of poly(dG) strands; AFM; CD spectroscopy;
D O I
10.1016/j.ab.2007.10.017
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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 0.1 M 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. (c) 2007 Elsevier Inc. All rights reserved.
引用
收藏
页码:71 / 78
页数:8
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