Identification and characterization of the guinea-pig cytomegalovirus glycoprotein H gene.
Subunit vaccines which target viral envelope glycoproteins offer promise for the prevention of congenital cytomegalovirus (CMV) infection. The guinea pig model of CMV infection is uniquely well suited to testing vaccines for prevention of congenital infection, since, in contrast to other animal cytomegaloviruses, the guinea pig CMV (GPCMV) crosses the placenta, producing intrauterine infection. Antibody to the CMV glycoproteins B (gB) and H (gH) appears to be important in conferring protective immunity. Unfortunately, little is known about specific GPCMV envelope glycoproteins. Sequencing of GPCMV genome fragments was therefore undertaken to test whether GPCMV encodes a gH homologue. Partial sequencing of the Hind III A fragment of the GPCMV genome revealed an open reading frame of 2,169 nucleotides capable of encoding a protein of 723 amino acids. Computer matrix analyses demonstrated identity between this ORF and the gH coding sequences of other herpesviruses. The GPCMV gH ORF encodes 12 highly conserved cysteine residues, contains 9 potential N-linked glycosylation sites, and has a predicted M(r) of 81.6 kDa. Northern blot hybridizations with gH-specific probes identified an abundant 5.1 kb mRNA with expression kinetics of an "early" gene. A polyclonal antiserum raised against a synthetic peptide derived from the deduced amino acid sequence of the gH ORF identified a virion-associated protein with an approximate M(r) of 85-kDa, the putative GPCMV gH, in immunoblot assays.
Brady
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Division of Infectious Diseases, Children's Hospital Research Foundation, Cincinnati, Ohio, USA.
Schleiss
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MR
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D002478:Cells, Cultured; D002874:Chromosome Mapping; D003587:Cytomegalovirus / Q000235:genetics*; D003586:Cytomegalovirus Infections / Q000235:genetics / Q000276:immunology / Q000517:prevention & control; D004279:DNA, Viral / Q000032:analysis / Q000235:genetics; D005347:Fibroblasts; D015870:Gene Expression; D005814:Genes, Viral; D016679:Genome, Viral; D006031:Glycosylation; D006168:Guinea Pigs; D015151:Immunoblotting; D066298:In Vitro Techniques; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010957:Plasmids; D012333:RNA, Messenger / Q000032:analysis / Q000378:metabolism; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D014759:Viral Envelope Proteins / Q000138:chemical synthesis / Q000235:genetics* / Q000276:immunology
D004279:DNA, Viral; D012333:RNA, Messenger; D012367:RNA, Viral; D014759:Viral Envelope Proteins; C071440:glycoprotein H, Cytomegalovirus