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

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

 


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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

Tsitologiya i Genetika 2008, vol. 42, no. 2, pp. 16-20

Institute of cell biology and genetic engineering, Zabolotnogo str. 148, Kyiv 03680, Ukraine



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, Tsitol Genet., 2008, vol. 42, no. 2, pp. 16-20.




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