title: Single-molecule imaging of DNA polymerase I (Klenow fragment) activity by atomic force microscopy.
abstract: We report a DNA origami-facilitated single-molecule platform that exploits atomic force microscopy to study DNA replication. We imaged several functional activities of the Klenow fragment of E. coli DNA polymerase I (KF) including binding, moving, and dissociation from the template DNA. Upon completion of these actions, a double-stranded DNA molecule was formed. Furthermore, the direction of KF activities was captured and then confirmed by shifting the KF binding sites on the template DNA.
authors.affiliation: Division of Physical Biology & Bioimaging Center, Shanghai Synchrotron Radiation Facility, CAS Key Laboratory of Interfacial Physics and Technology, Shanghai Institute of Applied Physics, Chinese Academy of Sciences, Shanghai 201800, China. libin@sinap.ac.cn fengzhang1978@hotmail.com.
authors.forename: J
authors.initials: J
authors.lastname: Chao
authors.forename: P
authors.initials: P
authors.lastname: Zhang
authors.forename: Q
authors.initials: Q
authors.lastname: Wang
authors.forename: N
authors.initials: N
authors.lastname: Wu
authors.forename: F
authors.initials: F
authors.lastname: Zhang
authors.forename: J
authors.initials: J
authors.lastname: Hu
authors.forename: C H
authors.initials: CH
authors.lastname: Fan
authors.forename: B
authors.initials: B
authors.lastname: Li
mesh_terms: D001665:Binding Sites; D002232:Carbocyanines / Q000737:chemistry; D004247:DNA / Q000737:chemistry*; D004256:DNA Polymerase I / Q000737:chemistry*; D004261:DNA Replication; D004277:DNA, Single-Stranded / Q000737:chemistry*; D004926:Escherichia coli / Q000201:enzymology*; D031541:Fluorescence Resonance Energy Transfer; D018625:Microscopy, Atomic Force; D053758:Nanoparticles / Q000737:chemistry; D036103:Nanotechnology
chemical_list: D002232:Carbocyanines; D004277:DNA, Single-Stranded; C089187:cyanine dye 3; C085321:cyanine dye 5; D004247:DNA; D004256:DNA Polymerase I