Expression from cloned DNA of biologically active glycoprotein C of herpes simplex virus type 1 in mammalian cells.
A DNA fragment of the herpes simplex virus type 1 genome encoding glycoprotein C (gC-1) has been cloned into different eukaryotic expression vectors for transient and stable expression of the glycoprotein in a number of cell lines. All of these expression vectors use a non-HSV promoter, such as the adenovirus major late promoter or murine leukemia virus long terminal repeat promoter to express gC-1 in COS and CHO cells or 3T3 cells. The gC-1 protein synthesized was fully glycosylated with both N- and O-linked oligosaccharides. Synthesis of the mature 120K gC-1 glycoprotein involved partially glycosylated 100K and 105K proteins and the non-glycosylated 70K protein as intermediate molecules. Immunofluorescence studies showed that the expressed gC-1 was localized intracellularly in the nuclear envelope as well as on the cell surface. The expressed gC-1 was biologically active and could act as a receptor for the complement component C3b in the absence of other HSV proteins.
Ghosh-Choudhury
N
N
McMaster University, Department of Biochemistry, Hamilton, Ontario, Canada.
Butcher
M
M
Ghosh
H P
HP
D000818:Animals; D002460:Cell Line; D003001:Cloning, Molecular; D004279:DNA, Viral / Q000235:genetics*; D005455:Fluorescent Antibody Technique; D015870:Gene Expression; D005814:Genes, Viral; D006031:Glycosylation; D008970:Molecular Weight; D010957:Plasmids; D015183:Restriction Mapping; D018139:Simplexvirus / Q000235:genetics*; D014162:Transfection; D014759:Viral Envelope Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification
D004279:DNA, Viral; D014759:Viral Envelope Proteins; C038535:glycoprotein gC, herpes simplex virus type 1