The α subunit of human chorionic gonadotropin hormone synthesized in insect cells using a baculovirus vector is biologically active

FEBS Letters

Hasnain, Seyed E.¹  Sridhar, Padma¹  Talwar, G.P.¹  Nakhai, Bita¹  Pal, Rahul¹ 

[1] National Institute of Immunology, New Delhi, 110067 India

关键词: Birth control vaccine;    Immunoreactivity;    Late promoter;    Post-translational modification;    Secretion;    Spodopiera frugiperda;    AcNPV;    Autographa californica nuclear polyhedrosis virus;    BEVS;    baculovirus expression vector system;    hCG;    human chorionic gonadotrophin;    ahCG;    gene coding for αhCG;    h;    hour(s);    moi;    multiplicity of infection;    pfu;    plaque forming unit;    pl;    post-infection;    Sf9;    Spodoptera frugiperda clone-9 insect cells;    wt;    wild-type;   

DOI  :  10.1016/0014-5793(91)80564-J

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

A recombinant baculovirus, vAcαhCG, 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 × 10⁴ 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 vAcαCG 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.

【 授权许可】

Unknown