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Journal of Virology, May 2005, p. 5315-5325, Vol. 79, No. 9
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.9.5315-5325.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Array Analysis of Simian Varicella Virus Gene Transcription in Productively Infected Cells in Tissue Culture

Steven B. Deitch,1 Donald H. Gilden,1,2 Mary Wellish,1 John Smith,1 Randall J. Cohrs,1* and Ravi Mahalingam1

Departments of Neurology,1 Microbiology, University of Colorado Health Sciences Center, Denver, Colorado 802622

Received 11 November 2004/ Accepted 21 December 2004

Simian varicella virus (SVV) is a neurotropic alphaherpesvirus of monkeys that is a model for varicella pathogenesis and latency. Like human varicella-zoster virus (VZV), SVV causes chicken pox (varicella), becomes latent in ganglia along the entire neuraxis, and reactivates to produce shingles (zoster). We developed macroarrays to determine the extent of viral transcription from all 70 predicted SVV open reading frames (ORFs) in infected cells in tissue culture. Cloned fragments (200 to 400 bp) from the 5' and 3' ends of each ORF were PCR amplified, quantitated, spotted onto nylon membranes, and fixed by UV cross-linking. Using a cDNA probe prepared from poly(A)+ RNA extracted from SVV-infected Vero cells at the height of the cytopathic effect (3 days after infection) and chemiluminescence for detection, transcripts corresponding to all SVV ORFs were identified. The abundance of each SVV transcript was compared with that previously demonstrated for VZV in infected tissue culture cells.

* Corresponding author. Mailing address: Department of Neurology, Mail Stop B182, 4200 E. 9th Avenue, Denver, CO 80262. Phone: (303) 315-8745. Fax: (303) 315-8720. E-mail: randall.cohrs{at}uchsc.edu.

Journal of Virology, May 2005, p. 5315-5325, Vol. 79, No. 9
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.9.5315-5325.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.





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