Atomic force microscopy of biochemically tagged DNA.
Small fragments of DNA of known length were made with the polymerase chain reaction. These fragments had biotin molecules covalently attached at their ends. They were subsequently labeled with a chimeric protein fusion between streptavidin and two immunoglobulin G-binding domains of staphylococcal protein A. This tetrameric species was expected to bind up to four DNA molecules via their attached biotin moieties. The DNA-protein complex was deposited on mica and imaged with an atomic force microscope. The images revealed the protein chimera at the expected location at the ends of the strands of DNA as well as the expected dimers, trimers, and tetramers of DNA bound to a single protein.
Murray
M N
MN
Human Genome Center, Lawrence Berkeley Laboratory, Berkeley, CA 94720.
Hansma
H G
HG
Bezanilla
M
M
Sano
T
T
Ogletree
D F
DF
Kolbe
W
W
Smith
C L
CL
Cantor
C R
CR
Spengler
S
S
Hansma
P K
PK
D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism; D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics / Q000648:ultrastructure*; D006801:Humans; D016252:Microscopy, Scanning Tunneling / Q000379:methods; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D012091:Repetitive Sequences, Nucleic Acid; D013205:Staphylococcal Protein A / Q000737:chemistry / Q000378:metabolism; D019809:Streptavidin
D001426:Bacterial Proteins; D011993:Recombinant Fusion Proteins; D013205:Staphylococcal Protein A; D004247:DNA; D019809:Streptavidin