Mapping and expression of a human cytomegalovirus major viral protein.
We constructed a DNA fragment map of low-passage Towne strain cytomegalovirus by analyzing cross-blot hybridization and hybridizations of isolated recombinant clones. The abundant late transcripts were located on this map by hybridization of labeled total RNA of virus-infected cells to blotted DNA fragments. The most abundant late transcript, carried by the 11.7-kilobase EcoRI fragment (EcoRI-G), was precisely mapped. The EcoRI fragment was fragmented and subcloned in a plasmid carrying simian virus 40 sequences (pSV-OH, constructed by Chi-Bom Chae, Department of Biochemistry, University of North Carolina, Chapel Hill). One resulting recombinant plasmid, pHD713SV2, was transferred to simian virus 40-transformed monkey kidney cells (COS-1) by DNA transfection. Synthesis of a cytomegalovirus-specific 67-kilodalton protein was detected in these cells by reaction of blotted proteins with virus-specific monoclonal antibody. The 67-kilodalton protein is a major phosphorylated protein found in virions; it is not glycosylated. The location of the gene for this 67-kilodalton protein is therefore assigned to the center of the L-unique region of human cytomegalovirus, at 0.37 to 0.39 map units.
D001483:Base Sequence; D002460:Cell Line; D003587:Cytomegalovirus / Q000235:genetics*; D004262:DNA Restriction Enzymes; D005796:Genes; D005814:Genes, Viral; D006801:Humans; D008970:Molecular Weight; D009693:Nucleic Acid Hybridization; D010957:Plasmids; D014176:Protein Biosynthesis; D014158:Transcription, Genetic; D014764:Viral Proteins / Q000235:genetics* / Q000302:isolation & purification
D014764:Viral Proteins; D004262:DNA Restriction Enzymes