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Russian Journal of Biotechnology
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High-Effective Production of Biologically Active Secreted Human Granulocyte Macrophage Colony-Stimulating Factor by Recombinant Vaccinia Virus
High-Effective Production of Biologically Active Secreted Human Granulocyte Macrophage Colony-Stimulating Factor by Recombinant Vaccinia VirusАвтор: A.A. Grazhdantseva, G.F. Sivolobova, A.V. Tkacheva, I.P. Gileva, E.V. Kuligina, V.A. Rikhter, and G.V. Kochneva Страница: 13-21
High-Effective Production of Biologically Active Secreted Human Granulocyte Macrophage Colony-Stimulating Factor by Recombinant Vaccinia Virus Biotekhnologiya, 2015, N 5, P. 13-21 UDC 577.21:578.28: 615.371 Section: “Producers, Biology, Selection, and Gene Engineering”
A.A. Grazhdantseva 1, 2, G.F. Sivolobova 1, 2, A.V. Tkacheva 1, 2, I.P. Gileva 1, E.V. Kuligina 2, V.A. Rikhter 2, and G.V. Kochneva 1, 2, * 1 The State Research Center for Virology and Biotechnology Vector, Novosibirskaya oblast, Kol'tsovo, 630559 Russia 2 The Institute for Chemical Biology and Fundamental Medicine, Russ. Acad. Sci., Siberian branch, Novosibirsk, 630090 Russia e-mail: g.v.kochneva@yandex.ru
A recombinant vaccinia virus strain VV-GMCSF-S1/3 that contains an insertion of a full-length DNA copy of messenger RNA of human granulocyte-macrophage colony-stimulating factor (GM-CSF) in the structural part of the viral thymidine kinase gene has been obtained. The expression of the GM-CSF gene as a part of the recombinant virus is under the control of the native vaccinia virus promoter р7.5K and results in the production of the mature form of the secreted protein with a molecular mass of 32 kDa. The biological activity of the GM-CSF was evaluated from the stimulation of the cytokine-dependent human erythroleukemia TF-1 cell proliferation. The level of the secretion of the biologically active human GM-CSF in the recombinant vaccinia virus / mammalian cells system was 1—40 μg/ml of culture medium. The recombinant strain of VV-GMCSF-S1/3 can be used as a producer of the glycosylated mature form of human GM-CSF, as well as a vector for the construction of oncolytic viruses and multivalent vaccine preparations.
Key words: cell proliferation assay XTT, human erythroleukemia TF-1 cells, GM-CSF, recombinant vaccinia virus, strain L-IVP.
20.05.2016, 1360 просмотров. |
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