New unstable variants of green fluorescent protein for studies of transient gene expression in bacteria.
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.
Andersen
J B
JB
Department of Microbiology, The Technical University of Denmark, DK-2800 Lyngby, Denmark.
Sternberg
C
C
Poulsen
L K
LK
Bjorn
S P
SP
Givskov
M
M
Molin
S
S
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
D017931:DNA Primers; D008164:Luminescent Proteins; D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins