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Appl Environ Microbiol. 1989 December; 55(12): 3119-3123
Copyright © 1989, American Society for Microbiology. All Rights Reserved.

High-Frequency Transformation, by Electroporation, of Lactococcus lactis subsp. cremoris Grown with Glycine in Osmotically Stabilized Media

Helge Holo{dagger},* and Ingolf F. Nes

1 Norwegian Dairies Association, Oslo, and Laboratory of Microbial Gene Technology, N-1432 Ås-NLH, 2 Norway

ABSTRACT

An efficient method for genetic transformation of lactococci by electroporation is presented. Highly competent lactococci for electrotransformation were obtained by growing cells in media containing high concentrations of glycine and 0.5 M sucrose as the osmotic stabilizers. These cells could be stored at –85°C without loss of competence. With Lactococcus lactis subsp. cremoris BC101, a transformation frequency of 5.7 x 107 transformants per µg of pIL253 DNA was obtained, which represents 5% of the surviving cells. All the lactococcal strains tested could be transformed by the present method.

FOOTNOTES

* Corresponding author.

{dagger} Present address: Laboratory of Microbial Gene Technology, P. O. Box 51, N-1432 Ås-NLH, Norway.

Appl Environ Microbiol. 1989 December; 55(12): 3119-3123
Copyright © 1989, American Society for Microbiology. All Rights Reserved.



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