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The α subunit of human chorionic gonadotropin hormone synthesized in insect cells using a baculovirus vector is biologically active
Author(s) -
Nakhai Bita,
Pal Rahul,
Sridhar Padma,
Talwar G.P.,
Hasnain Seyed E.
Publication year - 1991
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(91)80564-j
Subject(s) - biological activity , human chorionic gonadotropin , insect , vector (molecular biology) , protein subunit , hormone , biology , chemistry , endocrinology , biochemistry , recombinant dna , in vitro , botany , gene
A recombinant baculovirus, vAcα hCG , having a replacement of the viral polyhedrin gene with the cDNA encoding the α subunit of hCG was used to express αhCH, an extensively glycosylated hormone in insect cells. Virus‐infected cells, 72 h pi, secreted ∼ 11.3 μg αhCG/2 × 10 4 cells/ml which was identical to the native hormonal peptide in terms of electrophoretic mobility, immunoreactivity and bioactivity on association with β subunit, as evident by its binding to rat testicular cells and induction of steroidogenesis in a mouse Leydig cell bioassay system. The αhCG secreted into the medium represented ∼20–30% of the total hCG synthesized by vAcα CG infected insect cells. The implications of using a very late promoter, in a baculovirus expression system, for directing the transcription of a gene whose gene product requires extensive post‐translation modifications are discussed.