Jellyfish green fluorescent protein as a useful reporter for transient expression and stable transformation in Medicago sativa L.
The aim of the experiments reported herein was to transiently test different gene constructs using green fluorescent protein (GFP) as a reporter gene for a future localization of the maize beta-zein in the chloroplast of alfalfa ( Medicago sativa L.). The transient expression of two GFP genes was compared in alfalfa leaves to determine which of these two mutants is the easier to detect. Based on the intensity of fluorescence emitted, the GFP S65C gene was used to assemble a chloroplast-targeted GFP to verify the efficiency of the transit peptide for chloroplast targeting. A chloroplast-targeted fusion protein between beta-zein and GFP was then assembled, and this protein was observed to accumulate in small aggregates into the chloroplasts of transiently transformed cells. To the best of our knowledge, this is the first report of the GFP S65C gene being used to obtain transformed alfalfa plants expressing GFP.
Bellucci
M
M
Institute of Plant Genetics, Research Division of Perugia, CNR, Via Madonna Alta, 130, 06128 Perugia, Italy. michele.bellucci@igv.cnr.it
De Marchis
F
F
Mannucci
R
R
Arcioni
S
S
D002736:Chloroplasts / Q000235:genetics / Q000378:metabolism*; D018506:Gene Expression Regulation, Plant; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D000455:Medicago sativa / Q000166:cytology / Q000235:genetics / Q000378:metabolism*; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D009154:Mutation; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000166:cytology / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D015028:Zein / Q000235:genetics / Q000378:metabolism*
D008164:Luminescent Proteins; D010940:Plant Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D015028:Zein