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Restriction enzyme digestion patterns ofFrankia plasmids
Authors:P Simonet  P Normand  A Moiroud  M Lalonde
Institution:(1) Department des sciences forestières, Faculté de foresterie, Université Laval, G1K 7P4 Ste-Foy, Qué., Canada;(2) Laboratoire d'Ecologie Microbienne, ERA 848, Université Lyon 1, 69621 Villeurbanne, Cedex, France
Abstract:Summary After the initial screening of more than 200Frankia strains, the plasmid DNA observed in eight Frankiae was analyzed.In situ lysis was performed to obtain an estimate of their copy number and molecular weight. Four plasmid classes were distinguished, 7–9, 18–20, 30–35 and 50–55 kb. Twelve plasmids were thus analysed with restriction enzymes to determine their plasmid restriction patterns.While someFrankia plasmids with comparable molecular weights were found to be heterologous in their restriction enzyme pattern, an 8 kb plasmid found in bothFrankia sp. ArI3, isolated fromAlnus rubra andFrankia sp. CpI1 isolated fromComptonia peregrina showed undistinguishable lsquofingerprintsrsquo. Furthermore, an 18 kb plasmid found in the same two strains, also showed homologous restriction enzyme patterns. However, the copy numbers of the two ArI3 plasmids were higher than those of the CpI1 plasmids.Similarly, strains ACN1AG, 
$$TX41_{b^--- }^{AC}  and TX38_{b^--- }^{AC} $$
, isolated fromAlnus crispa all contained a 50 kb plasmid, and the three plasmids were found upon restriction analysis to be undistinguishable.In one strain, ARgX17c isolated fromAlnus rugosa, it was found through restriction enzyme analysis that two plasmids of a similar molecular weight were in fact heterologous.The possible origin of the homologous plasmids and their potential as specificFrankia markers to be used in ecological studies are discussed.
Keywords:Actinorhizae  Alnus  Frankia  Nitrogen fixation  Plasmids  Restriction analysis  Symbiosis
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