GFP and Family: Illuminating High Throughput Drug Screening

Published: 13 Feb 2006

By Marena Subitra Rajeevan, Research Analyst, Healthcare Division
marena.subitra@frost.com

Green fluorescent protein (GFP) was first isolated from the near colorless Pacific jellyfish, Aequorea victoria nearly three decades ago. The molecular cloning of GFP cDNA and its subsequent expression as a fully functional protein provided molecular and cellular biologist with an interesting and invaluable tool to study cellular processes. The uniqueness of GFP lies in its ability to auto-fluoresce upon expression with a suitable promoter. Unlike more conventional chemiluminescent, bioluminescent and fluorescent systems, GFP fluorescence does not require any external substrates or co-factors with the exception of molecular options. Additionally, GFP has been engineered to suit a variety of different applications and produce different colors such as yellow, blue and red.

The GFP Family and One-step Fluorescence

The green glow of GFP has enlightened scientists on protein localization, expression levels and protein-protein interactions in many complex cellular processes such as embryonic development. Something often difficult to achieve with more cytotoxic chemical reporters assays. Additionally, it has also had what can only be described as ‘fun’ applications, as scientists fascinated the public, with fluorescent glowing mice, plants, bunnies and most recently commercially available ‘glow-fish’.

Illuminating Technologies

GFP was initially utilized as a simple reporter gene. It excelled at this function due to its inherent ability to produce fluorescence when expressed with a suitable promoter in almost any cellular system. The key property of GFP that allowed it to advance past just qualitative applications was the quantitative nature of its expression. One unit of GFP fluorescence is equivalent to a single gene being expressed. Therefore any measurement of fluorescence levels indicate not only where and when the gene is expressed / functioning but also gives an indication of expression levels. The low cytotoxicity of GFP also lends it to be applied in live cell microscopy and to the monitoring of growth cultures in the fermentation industry.

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