The aim of this paper is to transiently test different gene constructs with 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 green fluorescent protein (GFP) genes was compared in alfalfa leaves to see 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. Then, a chloroplast-targeted fusion protein between beta-zein and GFP was assembled, and this protein accumulated in small aggregates into the chloroplasts of transiently transformed cells. Furthermore, the GFP S65C gene was used, for the first time to our knowledge, to obtain transformed alfalfa plants expressing GFP.
Jellyfish green fluorescent protein as a useful reporter in Medicago sativa L. for transient expression and stable transformation
Bellucci Michele;De Marchis Francesca;Arcioni Sergio
2003
Abstract
The aim of this paper is to transiently test different gene constructs with 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 green fluorescent protein (GFP) genes was compared in alfalfa leaves to see 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. Then, a chloroplast-targeted fusion protein between beta-zein and GFP was assembled, and this protein accumulated in small aggregates into the chloroplasts of transiently transformed cells. Furthermore, the GFP S65C gene was used, for the first time to our knowledge, to obtain transformed alfalfa plants expressing GFP.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
