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Expression of biologically active human granulocyte‐macrophage colony‐stimulating factor in the silkworm (Bombyx mori)
Author(s) -
Shi X.,
Qin J.,
Zhu J.,
Zhu D.
Publication year - 1996
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1111/j.1470-8744.1996.tb00402.x
Subject(s) - polyhedrin , bombyx mori , recombinant dna , microbiology and biotechnology , hemolymph , biology , bombycidae , nuclear polyhedrosis virus , isoelectric point , granulocyte macrophage colony stimulating factor , recombinant virus , baculoviridae , complementary dna , virus , virology , biochemistry , enzyme , spodoptera , gene , in vitro
Using virus derived from Bombyx mori (silkworm) nuclear polyhedrosis virus (BmNPV), we constructed an infectious recombinant virus carrying the human granulocyte‐macrophage colony‐stimulating factor (hGM‐CSF) cDNA placed downstream from the polyhedrin promoter. The BmN cells infected with the recombinant baculovirus BmNPV‐GM‐CSF expressed recombinant (r)hGM‐CSF up to 3.9 times 10(5) colony‐formation units/ml in the medium. The maximum activity of rhGM‐CSF was 4.3 times 10(6) colony‐formation units/ml in the haemolymph of silkworm (Bombyx mori) larvae infected with BmNPV‐GM‐CSF. Three distinct species of GM‐CSF of molecular masses 15, 18 and 20 kDa were detected by immunoblotting. The specific activity of rhGM‐CSF purified from the haemolymph was up to 2.92 times 10(7) colony‐formation units/mg. The isoelectric point of the purified rhGM‐CSF was 5.5‐5.7.