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J Virol, August 1998, p. 6832-6837, Vol. 72, No. 8
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Localization of Varicella-Zoster Virus Gene 21 Protein in Virus-Infected Cells in Culture

Ravi Mahalingam,1,* Robert Lasher,2 Mary Wellish,1 Randall J. Cohrs,1 and Donald H. Gilden1,3

Departments of Neurology,1 Cellular and Structural Biology,2 and Microbiology,3 University of Colorado Health Sciences Center, Denver, Colorado 80262

Received 16 October 1997/Accepted 4 May 1998

Although four varicella-zoster virus (VZV) genes have been shown to be transcribed in latently infected human ganglia, their role in the development and maintenance of latency is unknown. To study these VZV transcripts, we decided first to localize their expression products in productively infected cells. We began with VZV gene 21, whose open reading frame (ORF) is 3,113 bp. We cloned the 5' and 3' ends and the predicted antigenic segments of the ORF as 1292-, 1280-, and 880-bp DNA fragments, respectively, into the prokaryotic expression vector pGEX-2T. The three VZV 21 ORFs were expressed as approximately 75-, 73-, and 59-kDa glutathione S-transferase fusion proteins in Escherichia coli. To prepare polyclonal antibodies that would recognize all potential epitopes on the VZV gene 21 protein, rabbits were inoculated with a mixture of the three fusion proteins, and antisera were obtained and affinity purified. Immunohistochemical and immunoelectron microscopic analyses using these antibodies revealed VZV ORF 21 protein in both the nucleus and cytoplasm of VZV-infected cells. When these antibodies were applied to purified VZV nucleocapsids, intense staining was seen in their central cores.

* Corresponding author. Mailing address: Department of Neurology, Box B-182, University of Colorado Health Sciences Center, 4200 East 9th Ave., Denver, CO 80262. Phone: (303) 315-8100. Fax: (303) 315-8720. E-mail: ravi.mahalingam{at}uchsc.edu.

J Virol, August 1998, p. 6832-6837, Vol. 72, No. 8
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

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