Transformation of environmental Bacillus subtilis isolates by transiently inducing genetic competence.

Détails

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Etat: Public
Version: Final published version
ID Serval
serval:BIB_A8F59BA4A26D
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
Transformation of environmental Bacillus subtilis isolates by transiently inducing genetic competence.
Périodique
PLoS One
Auteur(s)
Nijland R., Burgess J.G., Errington J., Veening J.W.
ISSN
1932-6203 (Electronic)
ISSN-L
1932-6203
Statut éditorial
Publié
Date de publication
2010
Volume
5
Numéro
3
Pages
e9724
Langue
anglais
Résumé
Domesticated laboratory strains of Bacillus subtilis readily take up and integrate exogenous DNA. In contrast, "wild" ancestors or Bacillus strains recently isolated from the environment can only be genetically modified by phage transduction, electroporation or protoplast transformation. Such methods are laborious, have a variable yield or cannot efficiently be used to alter chromosomal DNA. A major disadvantage of using laboratory strains is that they have often lost, or do not display ecologically relevant physiologies such as the ability to form biofilms. Here we present a method that allows genetic transformation by natural competence in several environmental isolates of B. subtilis. Competence in these strains was established by expressing the B. subtilis competence transcription factor ComK from an IPTG-inducible promoter construct present on an unstable plasmid. This transiently activates expression of the genes required for DNA uptake and recombination in the host strain. After transformation, the comK encoding plasmid is lost easily because of its intrinsic instability and the transformed strain returns to its wild state. Using this method, we have successfully generated mutants and introduced foreign DNA into a number of environmental isolates and also B. subtilis strain NCIB3610, which is widely used to study biofilm formation. Application of the same method to strains of B. licheniformis was unsuccessful. The efficient and rapid approach described here may facilitate genetic studies in a wider array of environmental B. subtilis strains.
Mots-clé
Bacillus subtilis/genetics, Bacillus subtilis/metabolism, Bacterial Proteins/genetics, Bacterial Proteins/metabolism, Biofilms, DNA, Bacterial/genetics, Environmental Microbiology, Flow Cytometry/methods, Gene Expression Regulation, Bacterial, Genetic Techniques, Green Fluorescent Proteins/metabolism, Models, Genetic, Plasmids/metabolism, Promoter Regions, Genetic, Species Specificity, Transcription Factors/genetics, Transcription Factors/metabolism
Pubmed
Web of science
Open Access
Oui
Création de la notice
11/10/2016 16:29
Dernière modification de la notice
20/08/2019 16:13
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