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Production of Biologically Active Human Myelocytokines in Plants

A. S. Zvereva1*, L. E. Petrovskaya2, A. V. Rodina3, O. Y. Frolova1, P. A. Ivanov1, L. N. Shingarova2, T. V. Komarova4, Y. L. Dorokhov4, D. A. Dolgikh2, M. P. Kirpichnikov1,2, and J. G. Atabekov1

1Faculty of Biology, Lomonosov Moscow State University, 119991 Moscow, Russia; fax: (495) 938-0601; E-mail: zvereva@gmail.com; anna.zvereva@genebee.msu.ru

2Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, 117997 Moscow, Russia; fax: (495) 330-6983; E-mail: lpetr65@yahoo.com

3Moscow Institute of Medical Ecology, Simferopolsky Bulvar 8, 117638 Moscow, Russia; fax: (499) 113-4827; E-mail: allrodina@yandex.ru

4Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, 119991 Moscow, Russia; fax: (495) 938-3181; E-mail: dorokhov@genebee.msu.ru

* To whom correspondence should be addressed.

Received March 24, 2009; Revision received May 27, 2009
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.
KEY WORDS: granulocyte colony-stimulating factor, granulocyte macrophage colony-stimulating factor, viral vector, agroinjection

DOI: 10.1134/S0006297909110029