Escherichia coli cells penetrated by chrysotile fibers are transformed to antibiotic resistance by incorporation of exogenous plasmid DNA

N. Yoshida, K. Kodama, K. Nakata, Mitsuo Yamashita, T. Miwa

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

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 106 transformants/μg 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.

Original languageEnglish
Pages (from-to)461-468
Number of pages8
JournalApplied Microbiology and Biotechnology
Volume60
Issue number4
DOIs
Publication statusPublished - 2003
Externally publishedYes

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Serpentine Asbestos
Antibiotics
Escherichia coli
Plasmids
DNA
Anti-Bacterial Agents
Fibers
Asbestos
Agar
Positive ions
Cations
Electron microscopy
Drying
Membranes
Suspensions

ASJC Scopus subject areas

  • Biotechnology
  • Microbiology
  • Bioengineering

Cite this

Escherichia coli cells penetrated by chrysotile fibers are transformed to antibiotic resistance by incorporation of exogenous plasmid DNA. / Yoshida, N.; Kodama, K.; Nakata, K.; Yamashita, Mitsuo; Miwa, T.

In: Applied Microbiology and Biotechnology, Vol. 60, No. 4, 2003, p. 461-468.

Research output: Contribution to journalArticle

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