High efficiency transformation of Salmonella typhimurium and Salmonella typhi by electroporation |
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Authors: | David O''Callaghan and Alain Charbit |
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Affiliation: | (1) Unité de Programmation Moléculaire et Toxicologie Génétique, CNRS UA271, Inserm U163, Institut Pasteur, 25 rue du Dr. Roux, F-75105 Paris, France |
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Abstract: | Summary Salmonella typhimurium and S. typhi were transformd with high efficiency by electroporation. Transformation efficiencies of up to 1010 transformants per g of pBR322 were obtained. In contrast to chemical transformation methods, neither the smooth lipopolysaccharide of S. typhimurium nor the Vi capsular polysaccharide of S. typhi greatly affected transformation efficiency. The introduction of a galE mutation slightly improved transformation efficiency in S. typhimurium (< tenfold) while the Vi antigen of S. typhi had no detectable effect. The transformation efficiency of S. typhimurium with DNA derived from Escherichia coli was increased greatly by the removal of the hsd restriction system (100-fold). Under these conditions electroporation can be used for the routine and direct transformation of Salmonella strains with partially purified (alkaline lysis) plasmid DNA from E. coli. |
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Keywords: | Electroporation Salmonella typhimurium Salmonella typhit |
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