Green Fluorescent Protein as a Real Time Quantitative Reporter of Heterologous Protein Production
Citations Over TimeTop 10% of 1998 papers
Abstract
Since its cloning and commercial availability, applications of green fluorescent protein (GFP) as a reporter gene have become prevalent in many aspects of science. The attributes of GFP could also be applied to the area of heterologous protein production. The work described here represents the first experiments to use GFP as a generic tool to monitor protein production in bioprocess development. We have constructed a plasmid containing an operon fusion of the two reporter genes GFP and chloramphenicol acetyl transferase (CAT). CAT served as a "model" recombinant protein product to demonstrate the in situ quantifiable reporting mechanism of GFP. Our results indicate there is a direct correlation between the fluorescence intensity of GFP and the functional activity of the downstream CAT protein. In addition, there is a quantitative relationship between the level of CAT protein concentration and GFP fluorescence. These experiments provide the groundwork for using GFP as an in situ reporter gene for scale-up and process optimization of recombinant protein production.
Related Papers
- → Visualizing and quantifying protein secretion using aRenilla luciferase-GFP fusion protein(2000)19 cited
- → Green fluorescent protein is superior to blue fluorescent protein as a quantitative reporter of promoter activity in E. coli(2008)4 cited
- Expression and localization of s-lap/GFP fusion protein in Hela cell lines(2005)
- The expression of green fluorescent protein regulated by estrogen hormone in yeast cell(2006)
- Localization of overexpressed MDA-7/IL-24 protein(2006)