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Applied and Environmental Microbiology, January 2001, p. 293-299, Vol. 67, No. 1
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.1.293-299.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Evaluation of Biological and Physical Protection against Nuclease Degradation of Clay-Bound Plasmid DNA

Sandrine Demanèche,1 Lucile Jocteur-Monrozier,1 Hervé Quiquampoix,2 and Pascal Simonet1,*

Laboratoire d'Ecologie Microbienne, UMR 5557, Université Claude Bernard Lyon I, 69622 Villeurbanne Cedex,1 and Laboratoire de Science du Sol, INRA-ENSAM, 34060 Montpellier Cedex 01,2 France

Received 24 May 2000/Accepted 8 October 2000

In order to determine the mechanisms involved in the persistence of extracellular DNA in soils and to monitor whether bacterial transformation could occur in such an environment, we developed artificial models composed of plasmid DNA adsorbed on clay particles. We determined that clay-bound DNA submitted to an increasing range of nuclease concentrations was physically protected. The protection mechanism was mainly related to the adsorption of the nuclease on the clay mineral. The biological potential of the resulting DNA was monitored by transforming the naturally competent proteobacterium Acinetobacter sp. strain BD413, allowing us to demonstrate that adsorbed DNA was only partially available for transformation. This part of the clay-bound DNA which was available for bacteria, was also accessible to nucleases, while the remaining fraction escaped both transformation and degradation. Finally, transformation efficiency was related to the perpetuation mechanism, with homologous recombination being less sensitive to nucleases than autonomous replication, which requires intact molecules.

* Corresponding author. Mailing address: Laboratoire d'Ecologie Microbienne, UMR 5557, Université Lyon I, 43 bd du 11 Novembre 1918, 69622 Villeurbanne Cedex, France. Phone: 33 4 72 44 82 89. Fax: 33 4 72 43 12 23. E-mail: simonet{at}biomserv.univ-lyon1.fr.

Applied and Environmental Microbiology, January 2001, p. 293-299, Vol. 67, No. 1
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.1.293-299.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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