Staff Publications

Staff Publications

  • external user (warningwarning)
  • Log in as
  • language uk
  • About

    'Staff publications' is the digital repository of Wageningen University & Research

    'Staff publications' contains references to publications authored by Wageningen University staff from 1976 onward.

    Publications authored by the staff of the Research Institutes are available from 1995 onwards.

    Full text documents are added when available. The database is updated daily and currently holds about 240,000 items, of which 72,000 in open access.

    We have a manual that explains all the features 

Record number 423568
Title Random and cyclical deletion of large DNA segments in the genome of Pseudomonas putida
Author(s) Leprince, A.; Lorenzo, V. de; Völler, P.; Passel, M.W.J. van; Martins Dos Santos, V.A.P.
Source Environmental Microbiology 14 (2012)6. - ISSN 1462-2912 - p. 1444 - 1453.
DOI https://doi.org/10.1111/j.1462-2920.2012.02730.x
Department(s) Systems and Synthetic Biology
VLAG
WIMEK
Publication type Refereed Article in a scientific journal
Publication year 2012
Keyword(s) escherichia-coli genome - kt2440 - construction - sequence - system - genes - biodegradation - phenotypes - selection - insights
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
Comments
There are no comments yet. You can post the first one!
Post a comment
 
Please log in to use this service. Login as Wageningen University & Research user or guest user in upper right hand corner of this page.