Транзієнтна експресія GFP в культурі рослинних тканин in vitro за використання модульної системи вірусних векторів
Aim. Agrobacterium-mediated transient expression using viral-based vectors is one of the most effective method for recombinant protein production in plant. Transient expression of GFP using viral-based module system was studied in plant tissue culture in vitro. Methods. Regenerated shoots and callus clones of Nicotiana benthamiana were agroinfiltrated with viral-based module system. Protein extracts from GFP-positive tissues were resolved by non-reducing polyacrylamide gel electrophoresis. GFP from gel was eluted and GFP fluorescence was measured by fluorescence spectrometry with excitation filter at 395 nm and emission filter at 510 nm. Results. Regenerated shoots and callus clones lines accumulated GFP at 242.0±0.7 and 221.6±4.1 μg per g fresh tissue, respectively. The obtained level of transient expression is comparable with other plant production systems in early stage development. Conclusions. The developed technique shows promise for production of therapeutic proteins and antigens in the short term (14–16 days) by transient expression system in plant tissue culture in vitro.
Keywords: viral-based module system, transient expression, recombinant proteins, plant tissue culture, GFP.