Simultaneous synthesis of enzymatically active luciferase and biologically active β subunit of human chorionic gonadotropin in caterpillars infected with a recombinant baculovirus

FEBS Letters

Hasnain, Seyed E.¹  Sridhar, Padma¹  Jha, Prakash K.¹  Nakhai, Bita¹  Pal, Rahul¹ 

[1] National Institute of Immunology, Shahid Jeet Singh Marg, New Delhi-110067, India

关键词: β subunit of human chorionic gonadotropin;    βhCG;    Baculovirus system;    Dual expression;    Luciferase;    Recombinant DNA;    Secretion;    Glycosylation;    AcNPV;    Autographa californica nuclear polyhedrosis virus;    BEVS;    baculovirus expression vector system;    hCG;    human chorionic gonadotropin;    1gG-HRP;    immunoglobulin-horse radish peroxidase;    luc;    firefly luciferase;    MOI;    multiplicity of infection;    PBS;    phosphate-buffered saline;    p.i;    post infection;    Sf9;    Spodoptera frugiperda clone 9 cells;   

DOI  :  10.1016/0014-5793(92)81317-F

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The β subunit of human chorionic gonadotropin (βhCG), a secretory and extensively glycosylated hormone, and firefly luciferase, a non-secretory enzyme, were simultaneously synthesized in Spodoptera larvae upon infection with a dual expression recombinant baculovirus, vAcβhCG-luc. Luciferase was retained predominantly in the body tissue while βhCG was secreted into the hemolymph of infected larvae. Both the proteins were similar to their authentic counterparts in terms of immunoreactivity and bioactivity. The caterpillar-derived recombinant hCG exhibited reduced electrophoretic mobility on SDS-PAGE and increased biological activity as compared to the hCG expressed in insect cells in culture. The implications of using the larval system for expressing an extensively glycosylated protein are discusced.

【 授权许可】

Unknown