Applied Microbiology and Biotechnology

, Volume 52, Issue 4, pp 541–545

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

  • M. E. van der Rest
  • C. Lange
  • D. Molenaar
SHORT CONTRIBUTION

DOI: 10.1007/s002530051557

Cite this article as:
van der Rest, M., Lange, C. & Molenaar, D. Appl Microbiol Biotechnol (1999) 52: 541. doi:10.1007/s002530051557
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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.

Copyright information

© Springer-Verlag Berlin Heidelberg 1999

Authors and Affiliations

  • M. E. van der Rest
    • 1
  • C. Lange
    • 1
  • D. Molenaar
    • 1
  1. 1.Biotechnologisches Zentrallabor, Geb. 25.12, Heinrich-Heine-Universität, Universitätsstrasse 1, D-40225 Düsseldorf, Germany e-mail: molenaar@rz.uni-duesseldorf.de Tel.: +49-211-8111482 Fax: +49-211-8115370DE

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