Green fluorescent protein and its derivatives as versatile markers for gene expression in living Drosophila melanogaster, plant and mammalian cells

Jeffrey D. Plautz, Richard N. Day, Gina M. Dailey, Stephen B. Welsh, Jeffrey C. Hall, Shelley Halpain, Steve A. Kay

Research output: Contribution to journalArticle

99 Scopus citations

Abstract

We have investigated the utility of the green fluorescent protein (GFP) as a marker for gene expression in living adult Drosophila melanogaster (Dm) and cultured plant and mammalian cells. Using Dm, we generated transgenic flies bearing a glass-responsive gfp fusion gene to test the utility of GFP as a spatial reporter. In the adult living fly, GFP is clearly visible in the ocelli and the eye. We have optimized the use of filters for distinguishing the GFP signal from abundant autofluorescence in living Dm. In addition, we have used GFP to identify photoreceptor cells in pupal eye cultures that have been fixed and stained according to standard histological procedures, GFP was also detected in individual living plant cells following transient transfection of soybean suspension cultures, demonstrating that GFP is an effective transformation marker in plant cells. Similarly, transient transfection of mammalian cells with a modified form of GFP, S65T, allowed detection of single living cells expressing the reporter. This modified form of GFP gave a robust signal that was resistant to photobleaching. We then used a CellScan system exhaustive photon reassignment (EPR) deconvolution algorithm to generate high-resolution three-dimensional images of GFP fluorescence in the living cell.

Original languageEnglish (US)
Pages (from-to)83-87
Number of pages5
JournalGene
Volume173
Issue number1
DOIs
StatePublished - Jan 1 1996

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Keywords

  • Exhaustive photon reassignment
  • Fluorescence microscopy
  • GH3 cells
  • Glass gene
  • Image deconvolution
  • Particle bombardment
  • Pituitary
  • Soybean suspension culture
  • Transcription

ASJC Scopus subject areas

  • Genetics

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