Green-fluorescent protein as a new vital marker in plant cells
Article first published online: 5 MAR 2002
The Plant Journal
Volume 8, Issue 5, pages 777–784, November 1995
How to Cite
Sheen, J., Hwang, S., Niwa, Y., Kobayashi, H. and Galbraith, D. W. (1995), Green-fluorescent protein as a new vital marker in plant cells. The Plant Journal, 8: 777–784. doi: 10.1046/j.1365-313X.1995.08050777.x
- Issue published online: 5 MAR 2002
- Article first published online: 5 MAR 2002
- Received 27 June 1995; revised 4 August 1995; accepted 24 August 1995.
- Cited By
The green-fluorescent protein (GFP) from jellyfish Aequorea victoria has been used as a convenient new vital marker in various heterologous systems. However, it has been problematic to express GFP in higher eukaryotes, especially in higher plants. This paper reports that either a strong constitutive or a heat-shock promoter can direct the expression of GFP which is easily detectable in maize mesophyll protoplasts. In this single-cell system, bright green fluorescence emitted from GFP is visible when excited with UV or blue light even in the presence of blue fluorescence from the vacuole or the red chlorophyll autofluorescence from chloroplasts using a fluorescence microscope. No exogenous substrate, co-factor, or other gene product is required. GFP is very stable in plant cells and shows little photobleaching. Viable cells can be obtained after fluorescence-activated cell sorting based on GFP. The paper further reports that GFP can be detected in intact tissues after delivering the constructs into Arabidopsis leaf and root by microprojectile bombardment. The successful detection of GFP in plant cells relies on the use of a universal transcription enhancer from maize or the translation enhancer from tobacco mosaic virus (TMV) to boost the expression. This new reporter could be used to monitor gene expression, signal transduction, co-transfection, transformation, protein trafficking and localization, protein-protein interaction, cell separation and purification, and cell lineage in higher plants.