Expression of the glycoprotein H of murine cytomegalovirus and identification of an N-terminal antibody-binding region.
A series of overlapping fragments spanning the entire coding sequence of the gH gene of murine cytomegalovirus (MCMV) were expressed in Escherichia coli as fusion proteins with glutathione S-transferase (GST) using the pGEX expression system. A region of antibody-binding was mapped to the NH2-terminus of glycoprotein H (gH) between amino acid residues 26 and 90 on the basis of the reactivity of GST-gH fusion proteins with polyclonal antibodies to MCMV in Western blot analysis. Antibodies to gH were generated in mice immunized with the GST-gH fusion protein SK and shown to react with an 87-kDa polypeptide present in virion particles which was conserved in MCMV isolates obtained from diverse locations. They also recognized the gH protein in MCMV-infected cells, as well as gH expressed in Chinese Hamster Ovary cells. The antibodies to gH had a significant ELISA titer but no neutralizing activity in vitro. The antibody response to the GST-gH fusion protein did not modify the level of MCMV replication in the spleens of mice.
Xu
J
J
Department of Microbiology, University of Western Australia, Queen Elizabeth II Medical Centre, Nedlands.
Scalzo
A A
AA
Lyons
P A
PA
Shellam
G R
GR
D000818:Animals; D000914:Antibodies, Viral; D000956:Antigens, Viral / Q000032:analysis* / Q000276:immunology; D016466:CHO Cells; D002478:Cells, Cultured; D006224:Cricetinae; D003587:Cytomegalovirus / Q000737:chemistry*; D000939:Epitopes / Q000032:analysis; D004926:Escherichia coli / Q000235:genetics; D005260:Female; D006031:Glycosylation; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D011993:Recombinant Fusion Proteins / Q000032:analysis* / Q000276:immunology / Q000502:physiology; D013154:Spleen / Q000382:microbiology; D014611:Vaccination; D014759:Viral Envelope Proteins / Q000032:analysis* / Q000276:immunology / Q000502:physiology; D014771:Virion / Q000737:chemistry; D014779:Virus Replication
D000914:Antibodies, Viral; D000956:Antigens, Viral; D000939:Epitopes; D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins; C071440:glycoprotein H, Cytomegalovirus