title: Single-molecular AFM probing of specific DNA sequencing using RecA-promoted homologous pairing and strand exchange.
abstract: The specific sequence in a linearlized double-stranded DNA target has been identified at a single-molecular level by atomic force microscopy (AFM). This was accomplished using RecA-coated, single-stranded DNA probes which were paired with a specific complementary DNA sequence in a linear double-stranded DNA target by strand-exchange reaction at a homologous sequence site with target DNA. The sites of interaction between the nucleoprotein filaments and the double-stranded DNA targets were directly visualized by AFM in solution containing 4 mM magnesium acetate. Measurements of the position of RecA-coated probes paired to individual target DNA showed that DNA probes specifically paired at their corresponding homologous target sequences. Strand exchange promoted by RecA and the visualization by AFM provided a rapid and efficient way to identify homologous sequence on a single-molecule target DNA.
authors.affiliation: Department of Biological Information, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Yokohama, Japan.
authors.forename: G H
authors.initials: GH
authors.lastname: Seong
authors.forename: T
authors.initials: T
authors.lastname: Niimi
authors.forename: Y
authors.initials: Y
authors.lastname: Yanagida
authors.forename: E
authors.initials: E
authors.lastname: Kobatake
authors.forename: M
authors.initials: M
authors.lastname: Aizawa
mesh_terms: D001483:Base Sequence; D004247:DNA / Q000737:chemistry*; D015342:DNA Probes; D018625:Microscopy, Atomic Force / Q000379:methods*; D011938:Rec A Recombinases / Q000737:chemistry*
chemical_list: D015342:DNA Probes; D004247:DNA; D011938:Rec A Recombinases