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Natural and electroporation-mediated transformation of Methanococcus voltae protoplasts

Par Conseil national de recherches du Canada

AuteurRechercher : 1; Rechercher : 1; Rechercher : 1; Rechercher : 1
Affiliation
  1. Conseil national de recherches du Canada. Institut des sciences biologiques du CNRC
FormatTexte, Article
Résumé
The lack of high-efficiency transformation systems has severely impeded genetic research on methanogenic members of the kingdom Archaeobacteria. By using protoplasts of Methanococcus voltae and an integration vector, Mip1, previously shown to impart puromycin resistance, we obtained natural transformation frequencies that were about 80-fold higher (705 transformants per μg of transforming DNA) than that reported with whole cells. Electroporation-mediated transformation of M. voltae protoplasts with covalently closed circular Mip1 DNA was possible, but at lower frequencies of ca. 177 transformants per μg of vector DNA. However, a 380-fold improvement (3,417 transformants per μg of DNA) over the frequency of natural transformation with whole cells was achieved by electroporation of protoplasts with linearized DNA. This general approach, of using protoplasts, should allow the transformation of other methanogens, especially those that may be gently converted to protoplasts as a result of their tendency to lyse in hypotonic solutions.
Date de publication
Dans
  • Applied and Environmental Microbiology 60 (1994) : 903–907.
Langueanglais
Numéro du CNRCPATEL1994
NRCC 37372
Numéro NPARC9358454
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Identificateur de l’enregistrement819aed8c-1574-4049-9a27-2bb09f242b03
Enregistrement créé2009-07-10
Enregistrement modifié2020-04-27
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