Real-time single-molecule observation of rolling-circle DNA replication

被引：61
|
作者：
Tanner, Nathan A. [1 ]
Loparo, Joseph J. [1 ]
Hamdan, Samir M. [1 ]
Jergic, Slobodan [2 ]
Dixon, Nicholas E. [2 ]
van Oijen, Antoine M. [1 ]
机构：
[1] Harvard Univ, Sch Med, Dept Biol Chem & Mol Pharmacol, Boston, MA 02115 USA
[2] Univ Wollongong, Sch Chem, Wollongong, NSW 2522, Australia
来源：
NUCLEIC ACIDS RESEARCH | 2009年 / 37卷 / 04期
基金：
美国国家卫生研究院;
关键词：
POLYMERASE-III HOLOENZYME; PROCESSIVE REPLICATION; PROTEIN; STRAND; DYNAMICS; FORK; DENSITY; TARGETS; PRIMASE;
D O I：
10.1093/nar/gkp006
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
摘要：
We present a simple technique for visualizing replication of individual DNA molecules in real time. By attaching a rolling-circle substrate to a TIRF microscope-mounted flow chamber, we are able to monitor the progression of single-DNA synthesis events and accurately measure rates and processivities of single T7 and Escherichia coli replisomes as they replicate DNA. This method allows for rapid and precise characterization of the kinetics of DNA synthesis and the effects of replication inhibitors.
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页数：6
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