Atomic Force Microscopy of Biochemically Tagged DNA

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 90; no. 9; pp. 3811 - 3814
• Main Authors: Murray, Matthew N., Hansma, Helen G., Bezanilla, Magdalena, Sano, Takeshi, Ogletree, D. Frank, Kolbe, William, Smith, Casandra L., Cantor, Charles R., Spengler, Sylvia, Hansma, Paul K., Salmeron, Miquel
• Format: Journal Article
• Language: English
• Published: Washington, DC National Academy of Sciences of the United States of America 01.05.1993; National Acad Sciences; National Academy of Sciences
• Subjects: 550400 - Genetics; Analytical, structural and metabolic biochemistry; AZOLES; Bacterial Proteins - chemistry; Bacterial Proteins - metabolism; Base Sequence; BASIC BIOLOGICAL SCIENCES; BIOCHEMISTRY; Biological and medical sciences; BIOTIN; CARBOXYLIC ACIDS; CHEMISTRY; Chimeras; Cloning, Molecular; Deoxyribonucleic acid; DIMERS; DNA; DNA - genetics; DNA - ultrastructure; DNA POLYMERASES; DNA probes; ENZYMES; Fundamental and applied biological sciences. Psychology; General aspects, investigation methods; HETEROCYCLIC ACIDS; HETEROCYCLIC COMPOUNDS; Humans; Image resolution; Imaging; IMIDAZOLES; Mica; Microscopes; MICROSCOPY; Microscopy, Scanning Tunneling - methods; Molecular Sequence Data; Molecules; NUCLEIC ACIDS; NUCLEOTIDYLTRANSFERASES; ORGANIC ACIDS; ORGANIC COMPOUNDS; ORGANIC NITROGEN COMPOUNDS; ORGANIC SULFUR COMPOUNDS; PHOSPHORUS-GROUP TRANSFERASES; Physics; Polymerase Chain Reaction - methods; POLYMERASES; PROTEINS; Recombinant Fusion Proteins - chemistry; Recombinant Fusion Proteins - metabolism; Repetitive Sequences, Nucleic Acid; Scientific imaging; Sequencing; Staphylococcal Protein A - chemistry; Staphylococcal Protein A - metabolism; Streptavidin; TRANSFERASES; Trimers; VITAMIN B GROUP; VITAMINS; Proteins; Atomic force microscopy; Investigation method; DNA; Nucleotide fragment; Molecular complex; Molecular size; Chemical labelling; Biotine
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.90.9.3811

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 staphyloccocal 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.