Temporal nature of the promoter and not relative strength determines the expression of an extensively processed protein in a baculovirus system

FEBS Letters

Panda, Amulya K.²  Talwar, Gursaran P.³  Hasnain, Seyed E.¹  Sridhar, Padma¹  Pal, Rahul³ 

[1] Eukaryotic Gene Expression, National Institute of Immunology, Shahid Jit Singh Marg, New Delhi, 110067, India;Product Development,National Institute of Immunology, Shahid Jit Singh Marg, New Delhi, 110067, India;Immunoendocrinology Laboratories, National Institute of Immunology, Shahid Jit Singh Marg, New Delhi, 110067, India

关键词: Temporal promoter;    Secretion;    Biological activity;    Sialylation;    Insect cell;    AcNPV;    Autographa californien nuclear polyhedrosis virus;    hCG;    human chorionic gonadotropin;    Sf9;    Spodoptera frugiperda clone 9 insect cells;    βhCGPOL;    secretory recombinant βhCG synthesized under polyhedrin promoter control;    βhCGMP;    secretory recombinant βhCG synthesized under MP1 promoter control;    vAcβhCGPOL;    recombinant baeulovirus which expresses the βCG gene under polyhedrin promoter control;    vAcβhCGMP;    recombinant baculovirus which expresses the βhCG gene under MP1 promoter control.;   

DOI  :  10.1016/0014-5793(93)81179-4

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

We demonstrate that the expression of extensively modified and secreted heterologous proteins synthesized in the baculovirus expression vector system (BEVS) depends on the temporal nature of the promoter transcribing the foreign gene. The β subunit of the human chorionic gonadotropin, an extensively modified secretory glycoprotein hormone was expressed under the transcriptional control of the AcNPV basic protein gene promoter (MP) and the polyhedrin gene promoter (POL), respectively. MP, activated late in the infection cycle, is a weaker promoter when compared to the stronger very late POL promoter. Levels of secretion, immunoreactivity and bioactivity of recombinant proteins, βhCG_MP and βCG_POL. synthesized under control of the MP and POL promoter were compared. Secretion of βhCG_MP was relatively higher. Enzymatic analysis revealed that the synthesized protein was sialylated. Receptor binding assays and testosterone stimulation assays in a mouse Leydig cell system demonstrated that on a unit protein basis, βhCG_MP was biologically more active than βhCG_POL.

【 授权许可】

Unknown