VARICELLA-ZOSTER VIRUS (VZV) TRANSCRIPTION DURING LATENCY IN HUMAN GANGLIA - DETECTION OF TRANSCRIPTS MAPPING TO GENE-21, GENE-29, GENE-62,AND GENE-63 IN A CDNA LIBRARY ENRICHED FOR VZV RNA

Information on the extent of virus DNA transcription and translation i n infected tissue is crucial to an understanding of herpesvirus latenc y. To detect low-abundance latent varicella-zoster virus (VZV) transcr ipts, poly(A)(+) RNA extracted from latently infected human trigeminal ganglia was enriched for VZV transcripts by hybridization to biotinyl ated VZV DNA. After hybridization, the RNA-DNA hybrid was isolated by binding to avidin-coated beads and extensively washed, and the RNA was released by heat denaturation. A lambda-based cDNA library was then c onstructed from the enriched RNA. PCR and DNA sequencing of DNA extrac ted from the cDNA library revealed the presence of VZV genes 21, 29, 6 2, and 63, but not VZV genes 4, 10, 40, 51, and 61, in the enriched cD NA library. These findings confirm the detection of VZV gene 29 and 62 transcripts on Northern (RNA) blots prepared from latently infected h uman ganglia (J. L. Meier, R. P. Holman, K. D. Croen, J. E. Smialek, a nd S. E. Straus, Virology 193:193-200, 1993) and the presence of VZV g ene 21 transcripts in a cDNA library from mRNA of latently infected ga nglia (R J. Cohrs; K. Srock, M. B. Barbour, G. Owens, R Mahalingam, M. E. Devlin, M. Wellish, and D. H. Gilden, J. Virol. 68:7900-7908, 1994 ) and also reveal, for the first time, the presence of VZV gene 63 RNA in latently infected human ganglia.