Rapid Evaluation and Optimization of Recombinant Protein Production Using GFP Tagging

The isolation of recombinant proteins from bacterial or eukaryotic systems often requires a laborious optimization of expression and purification conditions. To greatly facilitate this procedure we included the green fluorescent protein (GFP) in bacterial expression vectors. This approach allowed us...

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Bibliographic Details
Published in:Protein expression and purification 2001-02, Vol.21 (1), p.220-223
Main Authors: Rücker, Elke, Schneider, Grit, Steinhäuser, Kim, Löwer, Roswitha, Hauber, Joachim, Stauber, Roland H.
Format: Article
Language:English
Subjects:
Cloning, Molecular
Escherichia coli - cytology
Escherichia coli - enzymology
Escherichia coli - growth & development
Genes, Reporter
GFP
Glutathione Transferase - genetics
Glutathione Transferase - isolation & purification
Glutathione Transferase - metabolism
Green Fluorescent Proteins
GST
Humans
Luminescent Proteins - analysis
Luminescent Proteins - isolation & purification
Microscopy, Fluorescence
Recombinant Fusion Proteins - biosynthesis
Recombinant Fusion Proteins - chemistry
Recombinant Fusion Proteins - isolation & purification
recombinant protein production
Transglutaminases - analysis
Transglutaminases - genetics
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Summary:The isolation of recombinant proteins from bacterial or eukaryotic systems often requires a laborious optimization of expression and purification conditions. To greatly facilitate this procedure we included the green fluorescent protein (GFP) in bacterial expression vectors. This approach allowed us to sensitively detect the GFP hybrid proteins already in intact bacterial cells using a fluorescence microscope. To rapidly analyze a variety of conditions essential for protein expression, the GFP signal, indicative of expression levels, was directly quantitated in live bacterial suspensions using a fluorescence plate reader. Thus, GFP tagging not only allows one to directly monitor protein expression in general but also appears to increase protein stability or solubility.
ISSN:1046-5928
1096-0279
DOI:10.1006/prep.2000.1373