Elaboration of an electroporation protocol for Bacillus cereus ATCC 14579

Authors: Turgeon, NathalieLaflamme, Christian; Ho, Jim; Duchaine, Caroline
Abstract: An electro-transformation procedure was established for Bacillus cereus ATCC 14579. Using early growth-stage culture and high electric field, the ectroporation efficiency was up to 2 x 10(9) cfu microg(-1) ml(-1) with pC194 plasmid DNA. The procedure was tested with three other plasmids, of various sizes, replication mechanisms and selection markers, and the transformation efficiencies ranged between 2 x 10(6) and 1 x 10(8) cfu microg(-1) ml(-)(1). The effects of two wall-weakening agents on electroporation rates were also evaluated. The transformation rate that was reached with our procedure is 10(3) times higher than that previously obtained with members of the Bacillus genus with similar plasmids, and 10(6) times superior than that achieved with available protocols for B. cereus. The proposed method is quick, simple, efficient with small rolling circle plasmids and large theta replicating plasmids with low copy number per cell, and suitable for many genetic manipulations that are not possible without high-efficiency transformation protocols.
Document Type: Article de recherche
Issue Date: 3 July 2006
Open Access Date: 25 August 2016
Document version: AM
Permalink: http://hdl.handle.net/20.500.11794/9408
This document was published in: Journal of microbiological methods, Vol. 67 (3), 543–548 (2006)
https://doi.org/10.1016/j.mimet.2006.05.005
Elsevier Biomedical
Alternative version: 10.1016/j.mimet.2006.05.005
16820234
Collection:Articles publiés dans des revues avec comité de lecture

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