A heat shock following electroporation induces highly efficient transformation of Corynebacterium glutamicum with xenogeneic plasmid DNA |
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| Authors: | M E van der Rest C Lange D Molenaar |
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| Institution: | Biotechnologisches Zentrallabor, Geb. 25.12, Heinrich-Heine-Universit?t, Universit?tsstrasse 1, D-40225 Düsseldorf, Germany e-mail: molenaar@rz.uni-duesseldorf.de Tel.: +49-211-8111482 Fax: +49-211-8115370, DE
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| Abstract: | 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.
Received: 26 March 1999 / Received revision: 9 June 1999 / Accepted: 11 June 1999 |
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