Mapping individual cosmid DNAs by direct AFM imaging.
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.
Allison
D P
DP
Health Sciences Research Division, Oak Ridge National Laboratory, Tennessee 37831-6123, USA. allisondp@ornl.gov
Kerper
P S
PS
Doktycz
M J
MJ
Thundat
T
T
Modrich
P
P
Larimer
F W
FW
Johnson
D K
DK
Hoyt
P R
PR
Mucenski
M L
ML
Warmack
R J
RJ
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
D004247:DNA