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Transformation of Bacillus subtilis by DNA bound on clay in non-sterile soil
Authors:E. Gallori  M. Bazzicalupo  L. Dal Canto  R. Fani  P. Nannipieri  C. Vettori  G. Stotzky
Affiliation:Department of Animal Biology and Genetics, University of Florence, Via Romana 17, 50125 Florence, Italy;Department of Soil Science, University of Florence, Florence, Italy;Laboratory of Microbial Ecology, Department of Biology, New York University, New York, USA
Abstract:Abstract Chromosomal DNA from Bacillus subtilis and different forms of plasmid pHV14 (covalently closed circular (CCC), linear monomer (M), and linear multimer (LM)) were adsorbed and bound on the clay mineral montmorillonite. After extensive washing of the clay-DNA complexes with DNA buffer (pH 7.5), approx. 25% of the chromosomal DNA, and approx. 30, 90, and 5%, respectively, of the CCC, M and LM form remained bound. Chromosomal and plasmid DNA bound on clay were capable of transforming competent cells, with different specific activities. The clay-DNA complexes persisted in non-sterile soil and retained transforming ability up to 15 days after their addition to the soil. DNA bound on montmortillonite was protected from the activity of Eco RI, supporting the evidence that DNA adsorbed on soil components was resistant to degradation by nucleases.
Keywords:Gene transfer    Natural transformation    Bacillus subtilis    Clay-DNA complex    Non-sterile soil
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