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Applied and Environmental Microbiology, September 1999, p. 3800-3804, Vol. 65, No. 9
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

An Effective Strategy, Applicable to Streptococcus salivarius and Related Bacteria, To Enhance or Confer Electroporation Competence

Nicole D. Buckley,1 Christian Vadeboncoeur,1 Donald J. LeBlanc,2,dagger Linda N. Lee,3 and Michel Frenette1,*

Groupe de Recherche en Écologie Buccale, Département de Biochimie et de Microbiologie, Faculté de Sciences et de Génie, et Faculté de Médecine Dentaire, Université Laval, Québec, Canada, G1K 1P41; Department of Oral Biology, Indiana University School of Dentistry, Indianapolis, Indiana 462022; and Department of Medicine, University of Texas Health Science Center at San Antonio, San Antonio, Texas 78284-77583

Received 2 March 1999/Accepted 10 June 1999

Despite the large number of techniques available for transformation of bacteria, certain species and strains are still resistant to introduction of foreign DNA. Some oral streptococci are among the organisms that can be particularly difficult to transform. We performed a series of experiments that involved manipulation of growth and recovery media and cell wall weakening, in which the electroporation conditions, cell concentration, and type and concentration of the transforming plasmid were varied. The variables were optimized such that a previously untransformable Streptococcus salivarius strain, ATCC 25975, could be transformed reproducibly at a level of 105 transformants per µg of DNA. The technique was used to introduce a plasmid into other strains of S. salivarius, including a fresh isolate. Moreover, the same technique was applied successfully to a wide range of oral streptococci and other gram-positive bacteria.

* Corresponding author. Mailing address: Groupe de Recherche en Écologie Buccale, Faculté de Médecine Dentaire, Université Laval, Québec, Canada G1K 7P4. Phone: (418) 656-2131, ext. 5502. Fax: (418) 656-2861. E-mail: Michel.Frenette{at}greb.ulaval.ca.

dagger Present address: Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, IN 46285.

Applied and Environmental Microbiology, September 1999, p. 3800-3804, Vol. 65, No. 9
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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