Baculovirus-mediated production of the human growth hormone in larvae of the silkworm, Bombyx mori.
To express the cDNA encoding human growth hormone (hGH) in larvae of Bombyx mori, B. mori nuclear polyhedrosis virus (BmNPV) was employed as an expression vector. For the construction of the recombinant virus, the hGH cDNA was inserted into the downstream of the strong polyhedrin promoter to achieve a high level expression. Immunoblot analysis revealed that the virus-mediated hGH was synthesized in the larvae and secreted into the hemolymph. The yield of the recombinant hGH synthesized in the larvae reached to a level of 160 micrograms/ml of hemolymph after purification. The purified recombinant hGH was confirmed to have both the same molecular weight and amino acid sequence at its N-terminal region as those of the natural counterpart. In addition, the biological activity of the recombinant hGH was comparable to that of the natural hGH in the growth-stimulating effect on rat Nb 2 Node lymphoma cells.
Kadono-Okuda
K
K
Laboratory of Biological Defense, National Institute of Sericultural and Entomological Science, Ibaraki, Japan.
Yamamoto
M
M
Higashino
Y
Y
Taniai
K
K
Kato
Y
Y
Chowdhury
S
S
Xu
J
J
Choi
S K
SK
Sugiyama
M
M
Nakashima
K
K
D000818:Animals; D012831:Bombyx / Q000254:growth & development / Q000378:metabolism*; D015246:Deoxyribonuclease EcoRI; D015252:Deoxyribonucleases, Type II Site-Specific; D005822:Genetic Vectors; D013006:Growth Hormone / Q000096:biosynthesis / Q000235:genetics*; D006458:Hemolymph / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting; D007700:Kinetics; D007814:Larva / Q000378:metabolism*; D017924:Nucleopolyhedroviruses / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000096:biosynthesis
D011994:Recombinant Proteins; D013006:Growth Hormone; D015246:Deoxyribonuclease EcoRI; D015252:Deoxyribonucleases, Type II Site-Specific; C029312:GGTACC-specific type II deoxyribonucleases