2.50
Hdl Handle:
http://hdl.handle.net/10033/240051
Title:
Random and cyclical deletion of large DNA segments in the genome of Pseudomonas putida.
Authors:
Leprince, Audrey; de Lorenzo, Víctor; Völler, Petra; van Passel, Mark W J; Martins dos Santos, Vitor A P
Abstract:
Cumulative site-directed mutagenesis is of limited suitability for the global analysis of the gene functions in the microbe's cellular network. In order to simplify and stabilize the genome of the soil bacterium Pseudomonas putida, we developed a recyclable three-step excision method based on the combination of customized mini-transposons and the FLP-FRT site-specific recombination system. To demonstrate the powerful potential of these tools, we first established insertion mutant libraries that allow users to study gene functions with respect either to phenotypic characteristics (single insertions) or to their involvement in predicted networks (double insertions). Based on these libraries, we generated as a proof-of-principle, single-deletion mutants lacking ~4.1% of the genome (~3.7% of the gene repertoire). A cyclical application of the method generated four double-deletion mutants of which a maximum of ~7.4% of the chromosome (~6.9% of the gene count) was excised. This procedure demonstrates a new strategy for rapid genome streamlining and gain of new insights into the molecular interactions and regulations.
Affiliation:
Systems and Synthetic Biology Group, Helmholtz-Centre for Infection Research, Braunschweig, Germany.
Citation:
Random and cyclical deletion of large DNA segments in the genome of Pseudomonas putida. 2012, 14 (6):1444-53 Environ. Microbiol.
Journal:
Environmental microbiology
Issue Date:
Jun-2012
URI:
http://hdl.handle.net/10033/240051
DOI:
10.1111/j.1462-2920.2012.02730.x
PubMed ID:
22429517
Type:
Article
Language:
en
ISSN:
1462-2920
Appears in Collections:
Publications of Dept. Gene Regulation and Differentiation (RDIF)

Full metadata record

DC FieldValue Language
dc.contributor.authorLeprince, Audreyen_GB
dc.contributor.authorde Lorenzo, Víctoren_GB
dc.contributor.authorVöller, Petraen_GB
dc.contributor.authorvan Passel, Mark W Jen_GB
dc.contributor.authorMartins dos Santos, Vitor A Pen_GB
dc.date.accessioned2012-08-27T12:34:58Z-
dc.date.available2012-08-27T12:34:58Z-
dc.date.issued2012-06-
dc.identifier.citationRandom and cyclical deletion of large DNA segments in the genome of Pseudomonas putida. 2012, 14 (6):1444-53 Environ. Microbiol.en_GB
dc.identifier.issn1462-2920-
dc.identifier.pmid22429517-
dc.identifier.doi10.1111/j.1462-2920.2012.02730.x-
dc.identifier.urihttp://hdl.handle.net/10033/240051-
dc.description.abstractCumulative site-directed mutagenesis is of limited suitability for the global analysis of the gene functions in the microbe's cellular network. In order to simplify and stabilize the genome of the soil bacterium Pseudomonas putida, we developed a recyclable three-step excision method based on the combination of customized mini-transposons and the FLP-FRT site-specific recombination system. To demonstrate the powerful potential of these tools, we first established insertion mutant libraries that allow users to study gene functions with respect either to phenotypic characteristics (single insertions) or to their involvement in predicted networks (double insertions). Based on these libraries, we generated as a proof-of-principle, single-deletion mutants lacking ~4.1% of the genome (~3.7% of the gene repertoire). A cyclical application of the method generated four double-deletion mutants of which a maximum of ~7.4% of the chromosome (~6.9% of the gene count) was excised. This procedure demonstrates a new strategy for rapid genome streamlining and gain of new insights into the molecular interactions and regulations.en_GB
dc.language.isoenen
dc.rightsArchived with thanks to Environmental microbiologyen_GB
dc.titleRandom and cyclical deletion of large DNA segments in the genome of Pseudomonas putida.en
dc.typeArticleen
dc.contributor.departmentSystems and Synthetic Biology Group, Helmholtz-Centre for Infection Research, Braunschweig, Germany.en_GB
dc.identifier.journalEnvironmental microbiologyen_GB
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