title: Rapid evaluation and optimization of recombinant protein production using GFP tagging.
abstract: 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.
authors.affiliation: Institute for Medical and Clinical Virology, University of Erlangen-Nürnberg, Schlossgarten 4, D-91054 Erlangen, Germany.
authors.forename: E
authors.initials: E
authors.lastname: Rücker
authors.forename: G
authors.initials: G
authors.lastname: Schneider
authors.forename: K
authors.initials: K
authors.lastname: Steinhäuser
authors.forename: R
authors.initials: R
authors.lastname: Löwer
authors.forename: J
authors.initials: J
authors.lastname: Hauber
authors.forename: R H
authors.initials: RH
authors.lastname: Stauber
mesh_terms: D003001:Cloning, Molecular; D004926:Escherichia coli / Q000166:cytology / Q000201:enzymology / Q000254:growth & development; D017930:Genes, Reporter; D005982:Glutathione Transferase / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D049452:Green Fluorescent Proteins; D006801:Humans; D008164:Luminescent Proteins / Q000032:analysis / Q000302:isolation & purification; D008856:Microscopy, Fluorescence; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification; D011503:Transglutaminases / Q000032:analysis / Q000235:genetics
chemical_list: D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D011503:Transglutaminases; D005982:Glutathione Transferase