Expression of biologically active human granulocyte-macrophage colony-stimulating factor in the silkworm (Bombyx mori).
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 x 10(5) colony-formation units/ml in the medium. The maximum activity of rhGM-CSF was 4.3 x 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 x 10(7) colony-formation units/mg. The isoelectric point of the purified rhGM-CSF was 5.5-5.7.
Shi
X
X
Department of Biochemistry, Nanjing University, People's Republic of China.
Qin
J
J
Zhu
J
J
Zhu
D
D
D000818:Animals; D012831:Bombyx; D002460:Cell Line; D004274:DNA, Recombinant / Q000235:genetics; D005822:Genetic Vectors; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000096:biosynthesis / Q000235:genetics*; D006801:Humans; D007814:Larva / Q000378:metabolism; D017924:Nucleopolyhedroviruses / Q000235:genetics*; D011994:Recombinant Proteins
D004274:DNA, Recombinant; D011994:Recombinant Proteins; D016178:Granulocyte-Macrophage Colony-Stimulating Factor