期刊
APPLIED BIOCHEMISTRY AND MICROBIOLOGY
卷 52, 期 7, 页码 685-691出版社
MAIK NAUKA/INTERPERIODICA/SPRINGER
DOI: 10.1134/S0003683816070036
关键词
human GM-CSF; erythroleukemia TF-1 cells; cell proliferation assay XTT; recombinant vaccinia virus; strain L-IVP
资金
- Ministry of Education of the Russian Federation [14.604.21.0057, RFMEFI60414X0057]
The recombinant vaccinia virus strain VV-GMCSF-S1/3, which contains an insertion of 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, was 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 NEuro7.5K; this results in the production of a mature form of the secreted protein with a molecular mass of 32 kDa. The biological activity of GM-CSF was evaluated by stimulation of the proliferation of cytokine-dependent human TF-1 erythroleukemia cells. The secretion level of biologically active human GM-CSF in the system of recombinant vaccinia virus/mammalian cells was 1-40 mu g/mL of culture medium. The recombinant strain 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.
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