New way to isolate simian virus 40 nucleoprotein complexes from infected cells: use of a thiol-specific reagent
Abstract
A new method for the isolation of simian virus 40 nucleoprotein complexes from nuclei of lytically infected cells is described. The method is based on the addition of a thiol-specific reagent, 5'5'-dithiobis(2-nitrobenzoic acid), to lysis and extraction buffers. By inhibiting an uncoating activity during simian virus 40 extraction, 5'5'-dithiobis (2-nitrobenzoic acid) allows the use of efficient extraction buffers, such as one containing Triton X-100 and EDTA, for the isolation of native simian virus 40 minichromosomes and virion-type structures. Use of the method is illustrated by following encapsidation of simian virus 40 minichromosomes in a pulse-chase experiment. Since 5'5'-dithiobis (2-nitrobenzoic acid) is an inhibitor of many different enzymes, the 5',5'-dithiobis (2-nitrobenzoic acid) extraction technique may be useful for the isolation of not only papovaviruses but also other viruses and possibly cellular chromatin.
Additional Information
© 1982 by the American Society for Microbiology. Received 17 September 1981/Accepted 12 November 1981. We thank Nancy Fritz for secretarial assistance. This work was supported by a grant to A.V. from the National Institutes of Health. F.M.B. was supported by a Massachusetts Institute of Technology Undergraduate Research Opportunity program. O.S. was supported by a National Science Foundation predoctoral fellowship. J.B. was supported by a departmental training grant from the National Institutes of Health.Attached Files
Published - BOYjvir82.pdf
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Additional details
- PMCID
- PMC256070
- Eprint ID
- 12426
- Resolver ID
- CaltechAUTHORS:BOYjvir82
- NIH
- Massachusetts Institute of Technology (MIT)
- NSF
- Created
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2008-11-25Created from EPrint's datestamp field
- Updated
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2021-11-08Created from EPrint's last_modified field