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

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. S...

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Autores principales:	Reindert Nijland, J Grant Burgess, Jeff Errington, Jan-Willem Veening
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2010
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/b3bf5060a89d48d6b24c70bac3551413
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spelling	oai:doaj.org-article:b3bf5060a89d48d6b24c70bac35514132021-11-25T06:25:24ZTransformation of environmental Bacillus subtilis isolates by transiently inducing genetic competence.1932-620310.1371/journal.pone.0009724https://doaj.org/article/b3bf5060a89d48d6b24c70bac35514132010-03-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/20300532/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203Domesticated 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.Reindert NijlandJ Grant BurgessJeff ErringtonJan-Willem VeeningPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 5, Iss 3, p e9724 (2010)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Reindert Nijland J Grant Burgess Jeff Errington Jan-Willem Veening Transformation of environmental Bacillus subtilis isolates by transiently inducing genetic competence.
description	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.
format	article
author	Reindert Nijland J Grant Burgess Jeff Errington Jan-Willem Veening
author_facet	Reindert Nijland J Grant Burgess Jeff Errington Jan-Willem Veening
author_sort	Reindert Nijland
title	Transformation of environmental Bacillus subtilis isolates by transiently inducing genetic competence.
title_short	Transformation of environmental Bacillus subtilis isolates by transiently inducing genetic competence.
title_full	Transformation of environmental Bacillus subtilis isolates by transiently inducing genetic competence.
title_fullStr	Transformation of environmental Bacillus subtilis isolates by transiently inducing genetic competence.
title_full_unstemmed	Transformation of environmental Bacillus subtilis isolates by transiently inducing genetic competence.
title_sort	transformation of environmental bacillus subtilis isolates by transiently inducing genetic competence.
publisher	Public Library of Science (PLoS)
publishDate	2010
url	https://doaj.org/article/b3bf5060a89d48d6b24c70bac3551413
work_keys_str_mv	AT reindertnijland transformationofenvironmentalbacillussubtilisisolatesbytransientlyinducinggeneticcompetence AT jgrantburgess transformationofenvironmentalbacillussubtilisisolatesbytransientlyinducinggeneticcompetence AT jefferrington transformationofenvironmentalbacillussubtilisisolatesbytransientlyinducinggeneticcompetence AT janwillemveening transformationofenvironmentalbacillussubtilisisolatesbytransientlyinducinggeneticcompetence
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Transformation of environmental Bacillus subtilis isolates by transiently inducing genetic competence.

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