Fusions between green fluorescent protein and beta-glucuronidase as sensitive and vital bifunctional reporters in plants

By fusing the genes encoding green fluorescent protein (GFP) and beta-glucuronidase (GUS) we have created a set of bifunctional reporter constructs which are optimized for use in transient and stable expression studies in plants. This approach makes it possible to combine the advantage of GUS, its h...

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Bibliographic Details
Published in:Plant molecular biology 1998-11, Vol.38 (5), p.861-873
Main Authors: Quaedvlieg, N.E.M. (Leiden Univ. (Netherlands). Inst. of Molecular Plant Sciences), Schlaman, H.R.M, Admiraal, P.C, Wijting, S.E, Stougaard, J, Spaink, H.P
Format: Article
Language:English
Subjects:
Arabidopsis - chemistry
Arabidopsis - genetics
ARABIDOPSIS THALIANA
Cloning, Molecular
EXPRESION GENICA
EXPRESSION DES GENES
Fluorescence
FORMATION DE NODOSITES
GENE EXPRESSION
Gene Expression Regulation, Plant
Genes, Reporter - genetics
Genes, Reporter - physiology
GENETIC TRANSFORMATION
Glucuronidase - genetics
Green Fluorescent Proteins
Immunoblotting
LOTUS
Lotus japonicus
Luminescent Proteins - genetics
MICROSCOPIA
MICROSCOPIE
MICROSCOPY
Microscopy, Confocal
Nicotiana tabacum
NODULACION
Plants - genetics
Plants, Genetically Modified
Plasmids - genetics
Potato virus X
Recombinant Fusion Proteins - analysis
Recombinant Fusion Proteins - genetics
ROOT NODULATION
Sensitivity and Specificity
TRANSFORMACION GENETICA
TRANSFORMATION GENETIQUE
Transformation, Genetic
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Summary:By fusing the genes encoding green fluorescent protein (GFP) and beta-glucuronidase (GUS) we have created a set of bifunctional reporter constructs which are optimized for use in transient and stable expression studies in plants. This approach makes it possible to combine the advantage of GUS, its high sensitivity in histochemical staining, with the advantages of GFP as a vital marker. The fusion proteins were functional in transient expression studies in tobacco using either DNA bombardment or potato virus X as a vector, and in stably transformed Arabidopsis thaliana and Lotus japonicus plants. The results show that high level of expression does not interfere with efficient stable transformation in A. thaliana and L. japonicus. Using confocal laser scanning microscopy we show that the fusion constructs are very suitable for promoter expression studies in all organs of living plants, including root nodules. The use of these reporter constructs in the model legume L. japonicus offers exciting new possibilities for the study of the root nodulation process.
ISSN:0167-4412
1573-5028
DOI:10.1023/A:1006182623165