Expression of biologically active human chorionic gonadotropin and its subunits by recombinant vaccinia virus.
Vaccinia virus (VV) expression vector was used to clone the genes for coding alpha and beta subunits of human chorionic gonadotropin (hCG). Recombinant viruses VSL3 and VSS1 containing these genes were selected as blue coloured plaques on the basis of co-expression of Escherichia coli beta-galactosidase in the infected cells. CV-1 cells when infected with VSL3 or VSS1 secreted 2.4 and 1.8 micrograms of alpha and beta hCG subunits, respectively, per 3 x 10(6) cells after 24 h of infection. The subunit proteins expressed individually had immunoreactivity with monoclonal and polyclonal antibodies specific to hCG. The subunit hormonal peptides associated with each other during co-infection to form the complete hCG dimer, which was biologically active as evident from the induction of steroidogenesis in a mouse Leydig cell system.
Chakrabarti
S
S
Gene Expression Laboratory, National Institute of Immunology, New Delhi, India.
Srinivasan
J
J
Lall
L
L
Rao
L V
LV
Talwar
G P
GP
D000818:Animals; D000911:Antibodies, Monoclonal; D002460:Cell Line; D006063:Chorionic Gonadotropin / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D018997:Chorionic Gonadotropin, beta Subunit, Human; D003001:Cloning, Molecular; D005814:Genes, Viral; D015292:Glycoprotein Hormones, alpha Subunit / Q000096:biosynthesis / Q000235:genetics; D006801:Humans; D051379:Mice; D010446:Peptide Fragments / Q000096:biosynthesis / Q000235:genetics; D010957:Plasmids; D011487:Protein Conformation; D011863:Radioimmunoassay; D014162:Transfection; D014616:Vaccinia virus / Q000235:genetics*; D010948:Viral Plaque Assay; D001616:beta-Galactosidase / Q000235:genetics
D000911:Antibodies, Monoclonal; D006063:Chorionic Gonadotropin; D018997:Chorionic Gonadotropin, beta Subunit, Human; D015292:Glycoprotein Hormones, alpha Subunit; D010446:Peptide Fragments; D001616:beta-Galactosidase