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Journal of Virology, November 2001, p. 11056-11070, Vol. 75, No. 22
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.22.11056-11070.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Structure and Assembly of Intracellular Mature Vaccinia Virus: Thin-Section Analyses

Gareth Griffiths,1,* Norbert Roos,2 Sybille Schleich,1 and Jacomine Krijnse Locker1

European Molecular Biology Laboratory, 69117 Heidelberg, Germany,1 and Electron Microscopy Unit for Biological Sciences, Department of Biology, University of Oslo, Blindern, N-0316 Oslo, Norway2

Received 23 March 2001/Accepted 21 August 2001

In the preceding study (see accompanying paper), we showed by a variety of different techniques that intracellular mature vaccinia virus (vaccinia IMV) is unexpectedly complex in its structural organization and that this complexity also extends to the underlying viral core, which is highly folded. With that analysis as a foundation, we now present different thin-section electron microscopy approaches for analyzing the IMV and the processes by which it is assembled in infected HeLa cells. We focus on conventional epoxy resin thin sections as well as cryosections to describe key intermediates in the assembly process. We took advantage of streptolysin O's ability to selectively permeabilize the plasma membrane of infected cells to improve membrane contrast, and we used antibodies against bone fide integral membrane proteins of the virus to unequivocally identify membrane profiles in thin sections. All of the images presented here can be rationalized with respect to the model put forward for the assembly of the IMV in the accompanying paper.

* Corresponding author. Mailing address: EMBL, Postfach 102209, 69117 Heidelberg, Germany. Phone: 49-6221-387267. Fax: 49-6221-387306. E-mail: griffiths{at}embl-heidelberg.de.

Journal of Virology, November 2001, p. 11056-11070, Vol. 75, No. 22
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.22.11056-11070.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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