title: Mapping individual cosmid DNAs by direct AFM imaging.
abstract: Individual cosmid clones have been restriction mapped by directly imaging, with the atomic force microscope (AFM), a mutant EcoRI endonuclease site-specifically bound to DNA. Images and data are presented that locate six restriction sites, predicted from gel electrophoresis, on a 35-kb cosmid isolated from mouse chromosome 7. Measured distances between endonuclease molecules bound to lambda DNA, when compared to known values, demonstrate the accuracy of AFM mapping to better than 1%. These results may be extended to identify other important site-specific protein-DNA interactions, such as transcription factor and mismatch repair enzyme binding, difficult to resolve by current techniques.
authors.affiliation: Health Sciences Research Division, Oak Ridge National Laboratory, Tennessee 37831-6123, USA. allisondp@ornl.gov
authors.forename: D P
authors.initials: DP
authors.lastname: Allison
authors.forename: P S
authors.initials: PS
authors.lastname: Kerper
authors.forename: M J
authors.initials: MJ
authors.lastname: Doktycz
authors.forename: T
authors.initials: T
authors.lastname: Thundat
authors.forename: P
authors.initials: P
authors.lastname: Modrich
authors.forename: F W
authors.initials: FW
authors.lastname: Larimer
authors.forename: D K
authors.initials: DK
authors.lastname: Johnson
authors.forename: P R
authors.initials: PR
authors.lastname: Hoyt
authors.forename: M L
authors.initials: ML
authors.lastname: Mucenski
authors.forename: R J
authors.initials: RJ
authors.lastname: Warmack
mesh_terms: D000818:Animals; D010582:Bacteriophage lambda / Q000235:genetics; D001665:Binding Sites / Q000235:genetics; D002874:Chromosome Mapping / Q000379:methods*; D003001:Cloning, Molecular; D003360:Cosmids / Q000235:genetics*; D004247:DNA / Q000235:genetics* / Q000378:metabolism; D007091:Image Processing, Computer-Assisted / Q000379:methods; D051379:Mice; D018625:Microscopy, Atomic Force / Q000379:methods*; D011485:Protein Binding
chemical_list: D004247:DNA