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A heat shock following electroporation induces highly efficient transformation of Corynebacterium glutamicum with xenogeneic plasmid DNA

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An improved method for the electrotransformation of wild-type Corynebacterium glutamicum (ATCC 13032) is described. The two crucial alterations to previously developed methods are: cultivation of cells used for electrotransformation at 18 °C instead of 30 °C, and application of a heat shock immediately following electrotransformation. Cells cultivated at sub optimal temperature have a 100-fold improved transformation efficiency (108 cfu μg−1) for syngeneic DNA (DNA isolated from the same species). A heat shock applied to these cells following electroporation improved the transformation efficiency for xenogeneic DNA (DNA isolated from a different species). In combination, low cultivation temperature and heat shock act synergistically and increased the transformation efficiency by four orders of magnitude to 2.5 × 106 cfu μg−1 xenogeneic DNA. The method was used to generate gene disruptions in C. glutamicum.

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Received: 26 March 1999 / Received revision: 9 June 1999 / Accepted: 11 June 1999

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van der Rest, M., Lange, C. & Molenaar, D. A heat shock following electroporation induces highly efficient transformation of Corynebacterium glutamicum with xenogeneic plasmid DNA. Appl Microbiol Biotechnol 52, 541–545 (1999). https://doi.org/10.1007/s002530051557

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  • DOI: https://doi.org/10.1007/s002530051557

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