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YOYO-1对DNA力学性质的影响。

Effect of YOYO-1 on the mechanical properties of DNA.

作者信息

Kundukad Binu, Yan Jie, Doyle Patrick S

机构信息

BioSystems and Micromechanics (BioSym) IRG, Singapore MIT Alliance for Research and Technology (SMART), Singapore.

出版信息

Soft Matter. 2014 Dec 28;10(48):9721-8. doi: 10.1039/c4sm02025a.

Abstract

YOYO-1 is a green fluorescent dye which is widely used to image single DNA molecules in solution for biophysical studies. However, the question of whether the intercalation of YOYO-1 affects the mechanical properties of DNA is still not clearly answered. Investigators have put forth contradicting data on the changes in persistence length of DNA. Here, we use atomic force microscopy to systematically study the changes in the mechanical properties of DNA due to the intercalation of YOYO-1. We first measured the persistence length, contour length and the bending angle distribution of the DNA-YOYO-1 complex. We find that the persistence length of DNA remains unaffected with the intercalation of YOYO-1. However the contour length increases linearly with about 38% increase at full saturation of 1 YOYO-1 per 4 base pairs of DNA. Next we measured the change in topology of relaxed closed circular DNA after the intercalation of YOYO-1. We find that YOYO-1 introduces supercoiling in closed circular DNA. Our observations indicate that the intercalation of YOYO-1 results in the underwinding of DNA duplex, but does not significantly change the persistence length.

摘要

YOYO-1是一种绿色荧光染料,广泛用于对溶液中的单个DNA分子进行成像,以开展生物物理研究。然而,YOYO-1的嵌入是否会影响DNA的机械性能这一问题仍未得到明确解答。研究人员针对DNA持久长度的变化给出了相互矛盾的数据。在此,我们使用原子力显微镜系统地研究了由于YOYO-1的嵌入而导致的DNA机械性能变化。我们首先测量了DNA-YOYO-1复合物的持久长度、轮廓长度和弯曲角度分布。我们发现,随着YOYO-1的嵌入,DNA的持久长度不受影响。然而,轮廓长度呈线性增加,在每4个DNA碱基对嵌入1个YOYO-1达到完全饱和时,轮廓长度增加约38%。接下来,我们测量了YOYO-1嵌入后松弛闭环DNA拓扑结构的变化。我们发现YOYO-1会在闭环DNA中引入超螺旋。我们的观察结果表明,YOYO-1的嵌入导致DNA双链解旋,但不会显著改变持久长度。

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