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An electron microscopic method for the mapping of proteins attached to nucleic acids

Wu, Madeline and Davidson, Norman (1978) An electron microscopic method for the mapping of proteins attached to nucleic acids. Nucleic Acids Research, 5 (8). pp. 2825-2846. ISSN 0305-1048.

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An electron microscopic method for demonstrating the presence of and mapping the positions of proteins specifically bound to nucleic acids is described. The nucleic acid-protein complex is treated with dinitro fluorobenzene under conditions such that dinitrophenyl (DNP) groups are attached to nucleophilic groups on the protein, with only a low level of random attachment to the nucleic acid. This product is treated with rabbit anti-DNP IgG. The position of the protein-(DNP)n(IgG)m complex on the nucleic acid strand can be observed by electron microscopy by protein free spreading methods and, in many cases, by cytochrome-c spread ing. If necessary for visualization by the latter method, the size of the labeled region can be increased by treatment with goat anti-rabbit IgG. High efficiency of electron microscopic labeling is achieved. Examples studied are: the adenovirus-2 DNA terminal protein, a protein covalently bound to 8V40 DNA, DNA polymerase I bound to DNA, E. coli RNA polymerase bound to T7 DNA and proteins UV cross linked to avian sarcoma virus RNA.

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Additional Information:Copyright © 1978 Oxford University Press. Received May 15, 1978. We are indebted to Dr. Klaus Bister for permission to quote preliminary results from our study with him of proteins crosslinked to ASV RNA by UV irradiation of virions. This research has been supported by grants GM 10991 and GM 20927 from the United States Public Health Service.
Issue or Number:8
Record Number:CaltechAUTHORS:WUMnar78a
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Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:4055
Deposited By: Tony Diaz
Deposited On:26 Jul 2006
Last Modified:02 Oct 2019 23:09

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