An effective strategy, applicable to Streptococcus salivarius and related bacteria, to enhance or confer electroporation competence.
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 10(5) transformants per microg 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.
Buckley
N D
ND
Groupe de Recherche en Ecologie Buccale, Département de Biochimie et de Microbiologie, Faculté de Sciences et de Génie, Université Laval, Québec, Canada, G1K 1P4.
Vadeboncoeur
C
C
LeBlanc
D J
DJ
Lee
L N
LN
Frenette
M
M
D015139:Blotting, Southern; D003470:Culture Media; D018274:Electroporation / Q000379:methods*; D005998:Glycine; D006094:Gram-Positive Bacteria / Q000235:genetics* / Q000254:growth & development; D010957:Plasmids / Q000235:genetics; D013291:Streptococcus / Q000235:genetics* / Q000254:growth & development; D014169:Transformation, Bacterial
D003470:Culture Media; D005998:Glycine