title: Green fluorescent protein as a real time quantitative reporter of heterologous protein production.
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.
authors.affiliation: Molecular and Cellular Biology Program and Department of Chemical and Biochemical Engineering, University of Maryland Baltimore County, Baltimore, Maryland 21250, USA.
authors.forename: C R
authors.initials: CR
authors.lastname: Albano
authors.forename: L
authors.initials: L
authors.lastname: Randers-Eichhorn
authors.forename: W E
authors.initials: WE
authors.lastname: Bentley
authors.forename: G
authors.initials: G
authors.lastname: Rao
mesh_terms: D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D003199:Computer Systems; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins; D016014:Linear Models; D008164:Luminescent Proteins / Q000235:genetics*; D009876:Operon; D010957:Plasmids / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics; D013050:Spectrometry, Fluorescence
chemical_list: D008164:Luminescent Proteins; D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins; D015500:Chloramphenicol O-Acetyltransferase