title: Procaryotic expression of phosphorylated tegument protein pp65 of human cytomegalovirus and application of recombinant peptides for immunoblot analyses.
abstract: The tegument of human cytomegalovirus (HCMV) contains a phosphorylated protein of 65 kilodaltons, termed pp65, which was reported to carry significant epitopes for the stimulation of the humoral immune response during natural infection. A monoclonal antibody directed against this protein was used to screen a lambda gt11 cDNA library for recombinant polypeptides. Two DNA fragments from purified lambda clones and one fragment from genomic DNA were used for cloning in a bacterial high-level expression vector. The resulting fusion proteins were tested for their reactivity with a panel of monoclonal antibodies directed against pp65 and with polyspecific anti-HCMV rabbit antisera. The binding site for all the monoclonal antibodies tested was found to be contained in one of the recombinant proteins with a viral portion of 26 amino acids. Immunoblot analyses with HCMV-positive human sera revealed that pp65 alone is not a reliable antigen for serodiagnosis but may be very useful in combination with other HCMV proteins.
authors.affiliation: Institut für Klinische und Molekulare Virologie, Universität Erlangen-Nürnberg, Federal Republic of Germany.
authors.forename: B
authors.initials: B
authors.lastname: Plachter
authors.forename: S
authors.initials: S
authors.lastname: Klages
authors.forename: S
authors.initials: S
authors.lastname: Hagelmann
authors.forename: W
authors.initials: W
authors.lastname: Britt
authors.forename: M P
authors.initials: MP
authors.lastname: Landini
authors.forename: G
authors.initials: G
authors.lastname: Jahn
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000096:biosynthesis; D015153:Blotting, Western; D002478:Cells, Cultured; D003587:Cytomegalovirus / Q000235:genetics*; D004274:DNA, Recombinant / Q000032:analysis; D015870:Gene Expression; D006801:Humans; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism*; D011817:Rabbits; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D015183:Restriction Mapping; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000276:immunology; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D004274:DNA, Recombinant; D010750:Phosphoproteins; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; D014764:Viral Proteins