A simple and rapid method for genetic transformation of lactic streptococci by electroporation

An electroporation procedure for the plasmid-mediated genetic transformation of intact cells of Streptococcus cremoris and Streptococcus lactis was performed. Ten different strains were transformed. The method was simple and rapid and yielded transformant colonies in 14 to 24 h. The method was optim...

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Bibliographic Details
Published in:Applied and Environmental Microbiology 1988-03, Vol.54 (3), p.655-660
Main Authors: POWELL, I. B, ACHEN, M. G, HILLIER, A. J, DAVIDSON, B. E
Format: Article
Language:English
Subjects:
analytical methods
Biological and medical sciences
Biotechnology
electric fields
Food industries
Food microbiology
Fundamental and applied biological sciences. Psychology
Genetic engineering
Genetic technics
genetic transformation
Genetics and Molecular Biology
membrane permeability
Methods. Procedures. Technologies
Milk and cheese industries. Ice creams
plasmide
plasmidios
plasmids
Streptococcus cremoris
Streptococcus lactis
technique analytique
tecnicas analiticas
transformacion genetica
transformation
transformation genetique
Vectors (cloning, transfer, expression). Insertion sequences and transposons
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Summary:An electroporation procedure for the plasmid-mediated genetic transformation of intact cells of Streptococcus cremoris and Streptococcus lactis was performed. Ten different strains were transformed. The method was simple and rapid and yielded transformant colonies in 14 to 24 h. The method was optimized for S. lactis LM0230, and transformation frequencies of between 1 X 10(4) and 5 X 10(5) transformants per microgram of purified plasmid (pMU1328) were achieved routinely. The optimized procedure involved lysozyme treatment of cells. Transformation of LM0230 occurred at comparable frequencies with PLS 1 (4.4 kilobase pair [kbp]), pMU 1328 (7.4 kbp), and pAM beta 1 (26.5 kbp). Plasmid DNA isolated from transformants had not undergone detectable deletions or rearrangements. Transformation was possible with plasmid DNA which was religated after restriction endonuclease digestion. Phage DNA-dependent transfection of S. lactis LM0230 and S. lactis C6 was also achieved.
ISSN:0099-2240
1098-5336
DOI:10.1128/aem.54.3.655-660.1988