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J Virol. 1988 April; 62(4): 1364-1372

Location, transcript analysis, and partial nucleotide sequence of the cytomegalovirus gene encoding an early DNA-binding protein with similarities to ICP8 of herpes simplex virus type 1.

Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.

ABSTRACT

The results presented here locate the gene encoding an early, nonvirion, single-stranded DNA-binding protein of human and simian strains of cytomegalovirus (CMV) [HCMV(Towne) DB140 and SCMV(Colburn) DB129, respectively] and provide additional evidence that this protein is the CMV homolog of the herpes simplex virus type 1 (HSV-1) major DNA-binding protein (ICP8), as proposed earlier (D. G. Anders, A. Irmiere, and W. Gibson, J. Virol. 58:253-262). The ICP8 gene was used as a probe in Southern analyses done at moderate stringency as an approach to locating similar sequences in the CMV genome. The BamHI K and EcoRI V fragments from the center of the long unique segment of HCMV(Towne) hybridized with the ICP8 probe and were in turn used to identify corresponding sequences in the EcoRI D fragment of SCMV(Colburn). RNA prepared from SCMV(Colburn)-infected cells directed the in vitro synthesis of DB129. If the RNA was first hybridized with the cloned 12.5-kilobase EcoRI D fragment, in vitro synthesis of DB129 was specifically inhibited. Additional hybrid-arrested in vitro translation experiments with subclones spanning the EcoRI D fragment demonstrated that the DB129 gene is located in the left half of that fragment, approximately bisected by a SalI site. RNA analyses identified 3.9-, 8.9-, and 10.0-kilobase RNA species expressed from this region. A partial nucleotide sequence of the Colburn region mapping within the boundaries of the 3.9-kilobase transcript, suspected to be the primary coding species, showed significant sequence similarity to the major DNA-binding protein gene homolog identified in B95-8 Epstein-Barr virus.

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