Use of the green fluorescent protein as a screenable marker for plant transformation

Abstract

A method for the production of transgenic plants is provided in which a vector carrying a gene encoding the green fluorescent protein is introduced into cells, the cells are screened for the protein and transformed cells are selected and regenerated. The cellular toxicity of the green fluorescent protein is circumvented by regulating expression of the gene encoding the protein or directing the protein to a subcellular compartment where it is not toxic to the cell. DNA constructs are provided for cell transformation in which the expression of a gene encoding the green fluorescent protein is placed under the control of an inducible promoter. In addition, DNA constructs are provided in which a nucleotide sequence encoding the green fluorescent protein is operably linked to a signal sequence which directs the expressed protein to a subcellular compartment where the protein is not toxic to the cell. Oxidative stress to plant cells transformed with GFP also can be ameliorated by transforming cells with an expression vector comprising genes encoding GFP and an oxygen scavenger enzyme such as superoxide dismutase. The toxicity of GFP in transformed plants can be eliminated by excising the screenable marker gene following detection of transformed cells or sectors. The FLP/FRT system is used in conjunction with GFP as a visible marker for transformation and FRT excision. A nucleotide sequence optimized for expression of the green fluorecent protein in plants is also provided.

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