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

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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^,* 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 lactococciby electroporation is presented. Highly competent lactococcifor electrotransformation were obtained by growing cells inmedia containing high concentrations of glycine and 0.5 M sucroseas the osmotic stabilizers. These cells could be stored at –85°Cwithout loss of competence. With Lactococcus lactis subsp. cremorisBC101, a transformation frequency of 5.7 x 10⁷ transformantsper µg of pIL253 DNA was obtained, which represents 5%of the surviving cells. All the lactococcal strains tested couldbe transformed by the present method.

FOOTNOTES

^* Corresponding author.

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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Martinussen, J., Schallert, J., Andersen, B., Hammer, K. (2001). The Pyrimidine Operon pyrRPB-carA from Lactococcus lactis. J. Bacteriol. 183: 2785-2794 [Abstract] [Full Text]
Émond, E., Lavallée, R., Drolet, G., Moineau, S., LaPointe, G. (2001). Molecular Characterization of a Theta Replication Plasmid and Its Use for Development of a Two-Component Food-Grade Cloning System for Lactococcus lactis. Appl. Environ. Microbiol. 67: 1700-1709 [Abstract] [Full Text]
Madsen, S. M., Mills, D., Djordjevic, G., Israelsen, H., Klaenhammer, T. R. (2001). Analysis of the Genetic Switch and Replication Region of a P335-Type Bacteriophage with an Obligate Lytic Lifestyle on Lactococcus lactis. Appl. Environ. Microbiol. 67: 1128-1139 [Abstract] [Full Text]
Østergaard, S., Brøndsted, L., Vogensen, F. K. (2001). Identification of a Replication Protein and Repeats Essential for DNA Replication of the Temperate Lactococcal Bacteriophage TP901-1. Appl. Environ. Microbiol. 67: 774-781 [Abstract] [Full Text]
Kraus, J., Geller, B. L. (2001). Cloning of Genomic DNA of Lactococcus lactis That Restores Phage Sensitivity to an Unusual Bacteriophage sk1-Resistant Mutant. Appl. Environ. Microbiol. 67: 791-798 [Abstract] [Full Text]
Walker, S. A., Klaenhammer, T. R. (2001). Leaky Lactococcus Cultures That Externalize Enzymes and Antigens Independently of Culture Lysis and Secretion and Export Pathways. Appl. Environ. Microbiol. 67: 251-259 [Abstract] [Full Text]
Dudley, E. G., Steele, J. L. (2001). Lactococcus lactis LM0230 contains a single aminotransferase involved in aspartate biosynthesis, which is essential for growth in milk. Microbiology 147: 215-224 [Abstract] [Full Text]
Garault, P., Letort, C., Juillard, V., Monnet, V. (2000). Branched-Chain Amino Acid Biosynthesis Is Essential for Optimal Growth of Streptococcus thermophilus in Milk. Appl. Environ. Microbiol. 66: 5128-5133 [Abstract] [Full Text]
Sinha, B., Francois, P., Que, Y.-A., Hussain, M., Heilmann, C., Moreillon, P., Lew, D., Krause, K.-H., Peters, G., Herrmann, M. (2000). Heterologously Expressed Staphylococcus aureus Fibronectin-Binding Proteins Are Sufficient for Invasion of Host Cells. Infect. Immun. 68: 6871-6878 [Abstract] [Full Text]
Goupil-Feuillerat, N., Corthier, G., Godon, J.-J., Ehrlich, S. D., Renault, P. (2000). Transcriptional and Translational Regulation of alpha -Acetolactate Decarboxylase of Lactococcus lactis subsp. lactis. J. Bacteriol. 182: 5399-5408 [Abstract] [Full Text]

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