title: New unstable variants of green fluorescent protein for studies of transient gene expression in bacteria.
abstract: Use of the green fluorescent protein (Gfp) from the jellyfish Aequorea victoria is a powerful method for nondestructive in situ monitoring, since expression of green fluorescence does not require any substrate addition. To expand the use of Gfp as a reporter protein, new variants have been constructed by the addition of short peptide sequences to the C-terminal end of intact Gfp. This rendered the Gfp susceptible to the action of indigenous housekeeping proteases, resulting in protein variants with half-lives ranging from 40 min to a few hours when synthesized in Escherichia coli and Pseudomonas putida. The new Gfp variants should be useful for in situ studies of temporal gene expression.
authors.affiliation: Department of Microbiology, The Technical University of Denmark, DK-2800 Lyngby, Denmark.
authors.forename: J B
authors.initials: JB
authors.lastname: Andersen
authors.forename: C
authors.initials: C
authors.lastname: Sternberg
authors.forename: L K
authors.initials: LK
authors.lastname: Poulsen
authors.forename: S P
authors.initials: SP
authors.lastname: Bjorn
authors.forename: M
authors.initials: M
authors.lastname: Givskov
authors.forename: S
authors.initials: S
authors.lastname: Molin
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D017930:Genes, Reporter; D014644:Genetic Variation; D049452:Green Fluorescent Proteins; D006207:Half-Life; D008164:Luminescent Proteins / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D016958:Pseudomonas putida / Q000235:genetics*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D037401:Scyphozoa / Q000235:genetics
chemical_list: D017931:DNA Primers; D008164:Luminescent Proteins; D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins