期刊
MOSCOW UNIVERSITY CHEMISTRY BULLETIN
卷 75, 期 5, 页码 280-285出版社
PLEIADES PUBLISHING INC
DOI: 10.3103/S0027131420050065
关键词
agroinfiltration; optimization of protein expression; design of experiment; transient expression; Nicotiana benthamiana
资金
- Blokhin National Medical Research Center of Oncology, Ministry of Health, Russia on the Development of technologies for the design, preparation, and application of personalized therapeutic antitumor vaccines [AAAA-A18-118032290146-5]
Optimizing the composition of an agroinfiltration solution is one way to boost the efficiency of the transient expression of the target protein in plants by usingAgrobacterium tumefaciens. We use the design of the experiment to investigate the effects that some chemical additives introduced into the agroinfiltration medium have on the yield of the target immunocytokine. It is shown that acetosyringone (at 600 mu M) and the surfactant Pluronic-68 (at 0.2%) markedly increase the yield of the target protein; lipoic acid (0-5 mu M) and povidone (0-0.5 g/L) do not have a significant effect on this parameter; and ascorbic acid at concentrations of 25-50 mM significantly lowers the yield. Using an agroinfiltration solution with the specific composition developed in this study, the yield of the target protein is found to be more than 500 mu g/g, which is sufficiently high for the practical production of recombinant proteins in plants.
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