Sindarovska Y.R., Sheludko Y.V., Gerasymenko I.M., Bannikova M.A., Kuchuk N.V.

Purification of recombinant GFP produced by Agrobacterum-mediated transient expression in Nicotiana excelsior

Cytology and Genetics 2008, v 42, number 2, 16-20

Green fluorescent protein (GFP) is commonly used as a reporter protein in a wide range of biological experiments. The efficient protocol of Agrobacterium-mediated transient expression in Nicotiana excelsior was applied for quick preparative production of recombinant GFP. The protein purification scheme has been developed and included ammonium sulfate precipitation and Q-sepharose anion-exchange chromatography. It results in obtaining of a fraction with about 85 % GFP homogeneity and the protein yield of about 75 %.

Key words: Nicotiana excelsior; Agrobacterium tumefaciens; transient expression; recombinant protein; GFP