title: Constitutive expression in human cells of herpes simplex virus type 1 glycoprotein B gene cloned in an episomal eukaryotic vector.
abstract: Expression of herpes simplex virus type 1 (HSV-1) glycoprotein B (gB-1) was obtained in human cells from the gB-1 gene cloned in the episomal replicating vector pBK-1, which contains the origin of replication and early region of the human papovavirus BK. Selective systems for the TK+ phenotype in TK-143B cells and for resistance to G418 in adenovirus 5-transformed 293 cells were used to obtain stable transformants that produced gB-1. While gB-1 expression in 143B cells required induction by HSV-1 early proteins, constitutive gB1 production was observed in 293 cells, where endogenous trans-acting factors probably replace the need for early viral products in the activation of the cloned gB-1 gene. The amount of recombinant gB-1 was comparable to that produced during HSV-1 lytic infection in human cells, due to amplification of the inserted gene in the replicating episomal vector. Expression of gB-1 was induced by cadmium and zinc when the promoter of the mouse metallothionein-I gene was placed upstream of gB1 structural sequences. The inducible system where the gB-1 gene is under the control of its own promoter could be employed to clarify the role of early viral products in induction of gB-1 synthesis. Constitutive expression of gB-1 in human cells could provide useful material for diagnostic purposes and for the preparation of a subunit vaccine against HSV infections.
authors.affiliation: Institute of Microbiology, University of Ferrara, Italy.
authors.forename: R
authors.initials: R
authors.lastname: Manservigi
authors.forename: R
authors.initials: R
authors.lastname: Gualandri
authors.forename: M
authors.initials: M
authors.lastname: Negrini
authors.forename: L
authors.initials: L
authors.lastname: Albonici
authors.forename: G
authors.initials: G
authors.lastname: Milanesi
authors.forename: E
authors.initials: E
authors.lastname: Cassai
authors.forename: G
authors.initials: G
authors.lastname: Barbanti-Brodano
mesh_terms: D015139:Blotting, Southern; D002461:Cell Line, Transformed; D003001:Cloning, Molecular; D004279:DNA, Viral / Q000032:analysis; D004797:Enzyme-Linked Immunosorbent Assay; D005784:Gene Amplification; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D006801:Humans; D010957:Plasmids; D011233:Precipitin Tests; D018139:Simplexvirus / Q000235:genetics* / Q000502:physiology; D014162:Transfection; D014759:Viral Envelope Proteins / Q000096:biosynthesis / Q000235:genetics*; D014779:Virus Replication
chemical_list: D004279:DNA, Viral; D014759:Viral Envelope Proteins; C040495:glycoprotein B, Simplexvirus