Escherichia coli cells penetrated by chrysotile fibers are transformed to antibiotic resistance by incorporation of exogenous plasmid DNA.
A suspension of recipient Escherichia coli cells in stationary phase, chrysotile asbestos, and pUC18 donor DNA spread over the surface of a Luria-Bertani agar plate using a streak bar several times, resulted in intracellular uptake of the plasmid DNA by the E. coli cells. The transformation efficiency was highest with a duration of cell exposure to chrysotile of more than 60 s and an agar concentration of 2%. To improve chrysotile-mediated transformation efficiency, we systematically optimized various conditions and parameters. In comparison to chrysotile exposure without cations, exposure with cations produced up to 100-fold more transformants. Optimized conditions resulted in 10(6) transformants/ micro g pUC18 DNA. The drastic physical change due to 'quick drying on the surface of the agar plate' when cells were exposed to chrysotile, was essential for chrysotile-mediated transformation. We suggest that DNA uptake mediated by chrysotile asbestos is the result of a mechanical physical transformation of E. coli, since the E. coli cells are not chemically competent. Electron microscopy of cells exposed to chrysotile suggested penetration of the E. coli membrane by chrysotile fibers. It is suggested that E. coli transformation by the plasmid DNA was the result of penetration by chrysotile fibers to which plasmid DNA is bound or adsorbed.
Yoshida
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N
Department of Biochemistry and Applied Biosciences, Miyazaki University, 1-1 Gakuen Kibanadai-Nishi, Japan. a04109u@cc.miyazaki-u.ac.jp
Kodama
K
K
Nakata
K
K
Yamashita
M
M
Miwa
T
T
D000362:Agar; D017632:Asbestos, Serpentine / Q000378:metabolism / Q000494:pharmacology*; D004351:Drug Resistance / Q000502:physiology*; D004926:Escherichia coli / Q000187:drug effects* / Q000235:genetics / Q000502:physiology; D008854:Microscopy, Electron; D010957:Plasmids / Q000235:genetics*; D014169:Transformation, Bacterial / Q000187:drug effects*; D001616:beta-Galactosidase / Q000378:metabolism
D017632:Asbestos, Serpentine; D000362:Agar; D001616:beta-Galactosidase