Production of biologically active human myelocytokines in plants.
An effective system for expression of human granulocyte and granulocyte macrophage colony-stimulating factors (hG-CSF and hGM-CSF) in Nicotiana benthamiana plants was developed using viral vector based on tobacco mosaic virus infecting cruciferous plants. The genes of target proteins were cloned into the viral vector driven by actin promoter of Arabidopsis thaliana. The expression vectors were delivered into plant cells by agroinjection. Maximal synthesis rate was detected 5 days after injection and was up to 500 and 300 mg per kg of fresh leaves for hG-CSF and hGM-CSF, respectively. The yield of purified hG-CSF and hGM-CSF was 100 and 50 mg/kg of fresh leaves, respectively. Recombinant plant-made hG-CSF and hGM-CSF stimulated proliferation of murine bone marrow and human erythroleucosis TF-1 cells, respectively, at the same rate as the commercial drugs.
Zvereva
A S
AS
Faculty of Biology, Lomonosov Moscow State University, Moscow, Russia. zvereva@gmail.com
Petrovskaya
L E
LE
Rodina
A V
AV
Frolova
O Y
OY
Ivanov
P A
PA
Shingarova
L N
LN
Komarova
T V
TV
Dorokhov
Y L
YL
Dolgikh
D A
DA
Kirpichnikov
M P
MP
Atabekov
J G
JG
D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology / Q000187:drug effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D015870:Gene Expression; D005818:Genetic Engineering / Q000379:methods*; D016179:Granulocyte Colony-Stimulating Factor / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology*; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology*; D006801:Humans; D051379:Mice; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology; D014026:Nicotiana / Q000235:genetics* / Q000378:metabolism
D011994:Recombinant Proteins; D016179:Granulocyte Colony-Stimulating Factor; D016178:Granulocyte-Macrophage Colony-Stimulating Factor