Faculty of Biology, Lomonosov Moscow State University, Moscow, Russia.
Biochemistry (Mosc). 2009 Nov;74(11):1187-94. doi: 10.1134/s0006297909110029.
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.
利用侵染十字花科植物的烟草花叶病毒为载体,构建了在本氏烟中高效表达人粒细胞集落刺激因子(hG-CSF)和人粒细胞-巨噬细胞集落刺激因子(hGM-CSF)的植物表达载体。目的蛋白基因克隆到由拟南芥肌动蛋白启动子驱动的病毒载体中,通过农杆菌介导的叶盘法转化本氏烟,5 天后在叶片中检测到目的蛋白的最大表达量,hG-CSF 和 hGM-CSF 的表达量分别达到 500 和 300 mg/kg 鲜重。从叶片中提取纯化 hG-CSF 和 hGM-CSF 的产量分别为 100 和 50 mg/kg 鲜重。重组植物来源的 hG-CSF 和 hGM-CSF 刺激小鼠骨髓和人红白血病 TF-1 细胞的增殖能力与商业化药物相当。