Single-molecule fluorescence studies on DNA looping

被引:13
|
作者
Jeong, Jiyoun [1 ]
Le, Tung T. [1 ]
Kim, Harold D. [1 ]
机构
[1] Georgia Inst Technol, Sch Phys, 837 State St, Atlanta, GA 30332 USA
基金
美国国家科学基金会; 美国国家卫生研究院;
关键词
DNA looping; J factor; Single-molecule fluorescence; FRET; PIFE; PERSISTENCE LENGTH; INTERSEGMENT TRANSFER; STRANDED-DNA; DOUBLE HELIX; IN-VITRO; PROTEINS; FLEXIBILITY; CYCLIZATION; DEPENDENCE; FRAGMENTS;
D O I
10.1016/j.ymeth.2016.04.005
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Structure and dynamics of DNA impact how the genetic code is processed and maintained. In addition to its biological importance, DNA has been utilized as building blocks of various nanomachines and nanostructures. Thus, understanding the physical properties of DNA is of fundamental importance to basic sciences and engineering applications. DNA can undergo various physical changes. Among them, DNA looping is unique in that it can bring two distal sites together, and thus can be used to mediate interactions over long distances. In this paper, we introduce a FRET-based experimental tool to study DNA looping at the single molecule level. We explain the connection between experimental measurables and a theoretical concept known as the J factor with the intent of raising awareness of subtle theoretical details that should be considered when drawing conclusions. We also explore DNA looping-assisted protein diffusion mechanism called intersegmental transfer using protein induced fluorescence enhancement (FIFE). We present some preliminary results and future outlooks. (C) 2016 Elsevier Inc. All rights reserved.
引用
收藏
页码:34 / 43
页数:10
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