Published in Scientific Bulletin. Series F. Biotechnologies, Vol. XXIV
Written by Oksana VARCHENKO1, Mykola KUCHUK, Myroslav PARII, Yuriy SYMONENKO
Transient gene expression in plants allows accumulation of large amount of recombinant proteins within a very short time. We optimized the protocol of transient green fluorescent protein (GFP) expression system in Aztec tobacco (Nicotiana rustica L.) plants. For transient transformation, the optimal optical density of the agrobacteria suspension for plant tissues infiltration was determined. The pSPV2303 genetic construct, contained the gfp gene from A. victoria under double 35S CaMV promoter, used for this purpose. The agrobacteria suspension was grown to OD = 1.0 initial optical density at wavelength 600 nm and the plants were infiltrated with the following final concentrations: 0.2; 0.4; 0.8; 1.0; 1.5; 2.0 without and with the addition of 4'-hydroxy-3',5'-dimethoxyacetophenone (acetosyringone) at 150 μM concentration. Spectrofluorometric and protein analysis were used for green fluorescent protein (GFP) detection. The highest level of expression obtained was using the OD = 1.0 agrobacteria optical density and there was not a detrimental effect on plant tissues.
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