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

Author:

Holo Helge1,Nes Ingolf F.1

Affiliation:

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 × 10 7 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.

Publisher

American Society for Microbiology

Subject

Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology

Reference18 articles.

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2. Transformation of bacteria by electroporation;Chassy B. M.;Trends Biotechnol.,1988

3. Construction of a vector plasmid family and its use for molecular cloning in Streptococcus lactis;Chopin A.;Biochimie,1988

4. High efficiency transformation of E. coli by high voltage electroporation;Dower W. J.;Nucleic Acids Res.,1988

5. Protoplast formation and regeneration in lactic streptococci;Fujita Y.;Agric. Biol. Chem.,1983

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