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An electron microscopic method for studying nucleic acid–protein complexes. Visualization of RNA polymerase bound to the DNA of bacteriophages T7 and T3
Author(s) -
Koller Th.,
Sogo J. M.,
Bujard H.
Publication year - 1974
Publication title -
biopolymers
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.556
H-Index - 125
eISSN - 1097-0282
pISSN - 0006-3525
DOI - 10.1002/bip.1974.360130514
Subject(s) - nucleic acid , chemistry , dna , ethidium bromide , polymerase , rna , biophysics , molecule , intercalation (chemistry) , nucleic acid thermodynamics , biochemistry , rna polymerase , microbiology and biotechnology , gene , organic chemistry , biology , base sequence
Double‐stranded DNA can be readily adsorbed on mica or directly on carbon coated grids from the surface of solutions containing ethidium bromide, actinomine, or propidium diiodide. The DNA molecules are unfolded, well separated, and show a length distribution similar to molecules prepared by protein monolayer techniques. Since the intercalating dyes tested do not lead to an increased apparent diameter of the nucleic acid the method is useful for the study of nucleic acid–protein complexes. As a model, the binding of E. coli RNA polymerase to phage T7 and T3 DNA was examined under different conditions. The enzyme can easily be identified and its position along the DNA molecule can be mapped.