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.
引用
收藏
页数：6
相关论文
共 50 条
  • [41] Single-Molecule Observation of DNA Replication Repair Pathways in E. coli
    Wollman, Adam J. M.
    Syeda, Aisha H.
    McGlynn, Peter
    Leake, Mark C.
    BIOPHYSICS OF INFECTION, 2016, 915 : 5 - 16
  • [42] Self-replication of circular DNA by a self-encoded DNA polymerase through rolling-circle replication and recombination
    Sakatani, Yoshihiro
    Yomo, Tetsuya
    Ichihashi, Norikazu
    SCIENTIFIC REPORTS, 2018, 8
  • [43] Of rolling-circle replication and conjugative transfer: variations on a protein-DNA interplay.
    不详
    PLASMID, 2005, 53 (01) : 27 - 27
  • [44] REAL-TIME OBSERVATION OF THE VIBRATION OF A SINGLE ADSORBED MOLECULE
    WATANABE, F
    MCCLELLAND, GM
    HEINZELMANN, H
    SCIENCE, 1993, 262 (5137) : 1244 - 1247
  • [45] Real-Time Imaging of Single-Molecule Enzyme Cascade Using a DNA Origami Raft
    Sun, Lele
    Gao, Yanjing
    Xu, Yan
    Chao, Jie
    Liu, Huajie
    Wang, Lianhui
    Li, Di
    Fan, Chunhai
    JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2017, 139 (48) : 17525 - 17532
  • [46] Real-time detection of human telomerase DNA synthesis by multiplexed single-molecule FRET
    Hentschel, Jendrik
    Badstubner, Mareike
    Choi, Junhong
    Bagshaw, Clive R.
    Lapointe, Christopher P.
    Wang, Jinfan
    Jansson, Linnea I.
    Puglisi, Joseph D.
    Stone, Michael D.
    BIOPHYSICAL JOURNAL, 2023, 122 (17) : 3447 - 3457
  • [47] Real-time observation of the dynamics of an individual rotaxane molecular shuttle using a single-molecule junction
    Chen, Sujun
    Su, Dingkai
    Jia, Chuancheng
    Li, Yanwei
    Li, Xingxing
    Guo, Xuefeng
    Leigh, David A.
    Zhang, Liang
    CHEM, 2022, 8 (01): : 243 - 252
  • [48] Real-time, single-molecule observation of biomolecular interactions inside nanophotonic zero mode waveguides
    Nemashkalo, A.
    Phipps, M. E.
    Hennelly, S. P.
    Goodwin, P. M.
    NANOTECHNOLOGY, 2022, 33 (16)
  • [49] INITIATION AND TERMINATION OF DNA-REPLICATION OF PLASMID PKYM VIA A ROLLING-CIRCLE MECHANISM
    YASUKAWA, H
    OZAKI, E
    MORITO, S
    MASAMUNE, Y
    JOURNAL OF GENERAL AND APPLIED MICROBIOLOGY, 1994, 40 (05): : 377 - 388
  • [50] Bacteriophage and host mutants causing the rolling-circle λ DNA replication early after infection
    Konopa, G
    Baranska, S
    Wegrzyn, A
    Wegrzyn, G
    FEBS LETTERS, 2000, 472 (2-3) : 217 - 220
12345