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PMID: 8483898 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

Atomic force microscopy of biochemically tagged DNA.

Murray MN, Hansma HG, Bezanilla M, Sano T, Ogletree DF, Kolbe W, Smith CL, Cantor CR, Spengler S, Hansma PK

Abstract

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.

MeSH Terms
Bacterial Proteins/chemistry,metabolism Base Sequence Cloning, Molecular DNA/genetics,ultrastructure Humans Microscopy, Scanning Tunneling/methods Molecular Sequence Data Polymerase Chain Reaction/methods Recombinant Fusion Proteins/chemistry,metabolism Repetitive Sequences, Nucleic Acid Staphylococcal Protein A/chemistry,metabolism Streptavidin
Chemicals
Bacterial Proteins Recombinant Fusion Proteins Staphylococcal Protein A DNA Streptavidin
Authors & Affiliations
10 authors, click to expand affiliations / ORCID
Murray M N
Human Genome Center, Lawrence Berkeley Laboratory, Berkeley, CA 94720.
Hansma H G
Bezanilla M
Sano T
Ogletree D F
Kolbe W
Smith C L
Cantor C R
Spengler S
Hansma P K
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
Published
1993-05-01
Pages
3811-4
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC46395
Subset
IM
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