New Unstable Variants of Green Fluorescent Protein for Studies of Transient Gene Expression in Bacteria
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| Published in: | Applied and Environmental Microbiology Vol. 64; no. 6; pp. 2240 - 2246 |
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| Main Authors: | , , , , , |
| Format: | Journal Article |
| Language: | English |
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01-06-1998
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| AbstractList | 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. Classifications Services AEM Citing Articles Google Scholar PubMed Related Content Social Bookmarking CiteULike Delicious Digg Facebook Google+ Mendeley Reddit StumbleUpon Twitter current issue Spotlights in the Current Issue AEM About AEM Subscribers Authors Reviewers Advertisers Inquiries from the Press Permissions & Commercial Reprints ASM Journals Public Access Policy AEM RSS Feeds 1752 N Street N.W. • Washington DC 20036 202.737.3600 • 202.942.9355 fax • journals@asmusa.org Print ISSN: 0099-2240 Online ISSN: 1098-5336 Copyright © 2014 by the American Society for Microbiology. For an alternate route to AEM .asm.org, visit: AEM 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. 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. |
| Author | Søren Molin Lars Kongsbak Poulsen Sara Petersen Bjørn Jens Bo Andersen Michael Givskov Claus Sternberg |
| AuthorAffiliation | Department of Microbiology, The Technical University of Denmark, DK-2800 Lyngby, 1 Microbial Discovery, Novo Nordisk A/S, DK-2880 Bagsværd, 2 and Bio Image, Novo Nordisk A/S, Mørkhøj Bygade 28, DK-2860 Søborg, 3 Denmark |
| AuthorAffiliation_xml | – name: Department of Microbiology, The Technical University of Denmark, DK-2800 Lyngby, 1 Microbial Discovery, Novo Nordisk A/S, DK-2880 Bagsværd, 2 and Bio Image, Novo Nordisk A/S, Mørkhøj Bygade 28, DK-2860 Søborg, 3 Denmark |
| Author_xml | – sequence: 1 givenname: J. B surname: ANDERSEN fullname: ANDERSEN, J. B organization: Department of Microbiology, The Technical University of Denmark, 2800 Lyngby, Denmark – sequence: 2 givenname: C surname: STERNBERG fullname: STERNBERG, C organization: Department of Microbiology, The Technical University of Denmark, 2800 Lyngby, Denmark – sequence: 3 givenname: L. K surname: POULSEN fullname: POULSEN, L. K organization: Microbial Discovery, Novo Nordisk A/S, 2880 Bagsværd, Denmark – sequence: 4 givenname: S. P surname: BJØRN fullname: BJØRN, S. P organization: Bio Image, Novo Nordisk A/S, Mørkhøj Bygade 28, 2860 Søbolg, Denmark – sequence: 5 givenname: M surname: GIVSKOV fullname: GIVSKOV, M organization: Department of Microbiology, The Technical University of Denmark, 2800 Lyngby, Denmark – sequence: 6 givenname: S surname: MOLINI fullname: MOLINI, S organization: Department of Microbiology, The Technical University of Denmark, 2800 Lyngby, Denmark |
| BackLink | http://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=2297276$$DView record in Pascal Francis https://www.ncbi.nlm.nih.gov/pubmed/9603842$$D View this record in MEDLINE/PubMed |
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| Issue | 6 |
| Keywords | Pseudomonadales Stability C terminal peptide Escherichia coli Gene expression Plasmid Coelenterata Reporter gene Cnidaria Green fluorescent protein Bacteria Pseudomonadaceae Mutation Recombinant protein Invertebrata Pseudomonas putida Enterobacteriaceae Half life |
| Language | English |
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| Notes | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 Corresponding author. Mailing address: Department of Microbiology, Building 301, The Technical University of Denmark, DK-2800 Lyngby, Denmark. Phone: 45 45 25 25 13. Fax: 45 45 88 73 28. E-mail: sm@im.dtu.dk. |
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| PublicationTitle | Applied and Environmental Microbiology |
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| SubjectTerms | Aequorea victoria Amino Acid Sequence Animals Bacteria Base Sequence Biological and medical sciences Biotechnology DNA Primers - genetics Escherichia coli Escherichia coli - genetics Fluorescence Fundamental and applied biological sciences. Psychology Gene Expression Genes Genes, Reporter Genetic Variation Green Fluorescent Proteins Half-Life Luminescent Proteins - genetics Luminescent Proteins - metabolism Methods Methods. Procedures. Technologies Microbiology Molecular Sequence Data Protein engineering Proteins Pseudomonas putida - genetics Psuedomonas putida Recombinant Proteins - genetics Recombinant Proteins - metabolism Scyphozoa - genetics |
| Title | New Unstable Variants of Green Fluorescent Protein for Studies of Transient Gene Expression in Bacteria |
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